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  • 1990-1994  (4,912)
  • 1955-1959  (2,468)
  • Organic Chemistry  (6,983)
  • gene expression
  • transformation
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Year
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  • 1
    Electronic Resource
    Electronic Resource
    Plasmid mediated silver resistance in Acinetobacter baumannii (1994)
    Springer
    BioMetals 7 (1994), S. 49-56 
    Details
    ISSN: 1572-8773
    Keywords: Acinetobacter ; conjugation ; curing ; plasmid ; silver uptake ; silver resistance ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Acinetobacter baumannii BL88, an environmental isolate, was resistant to 13 metals and 10 antibiotics. Plumbagin cured resistance to silver, cadmium, antimony, streptomycin and ampicillin at varying frequencies. However, only silver resistance transferred (1 × 10−6 recepient−1) to Escherichia coli K12 during conjugation. Correspondingly there was transfer of a 54 kb plasmid (pUPI199) from A. baumannii BL88. The plasmid transformed E. coli DH5α cells at a frequency of 1 × 10−8 recepient−1. The growth rate of E. coli DH5; (pUPI199) was slower as compared with E. coli DH5α. Plasmid pUPI199 was 76 and 9.6% stable in the host A. baumannii BL88 in the presence and absence of selection pressure, respectively. A. baumannii BL88 was found to accumulate and retain silver whereas E. coli DH5α (pUPI199) effluxed 63% of the accumulated silver ions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00205194
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  • 2
    Electronic Resource
    Electronic Resource
    Canonical correlation and reduction of multiple time series (1994)
    Springer
    Annals of the Institute of Statistical Mathematics 46 (1994), S. 625-631 
    Details
    ISSN: 1572-9052
    Keywords: Rank of multiple time series ; transformation ; spectrum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Mathematics
    Notes: Abstract It is shown that a degenerate rankd-variate stationary time series can be reduced to a full rank time series of lower dimension via an orthogonal transformationT provided that ρ, the canonical correlation between past and future of the time series is strictly less than one. Procedures for estimation of rank of the multiple time series,T and testing ρ=1 are outlined, the latter is related to testing the unit root hypothesis in ARMA models.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00773471
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  • 3
    Electronic Resource
    Electronic Resource
    Effect of radical scavengers on TNFα-mediated activation of the uPA in cultured cells (1994)
    Springer
    Cellular and molecular life sciences 50 (1994), S. 958-962 
    Details
    ISSN: 1420-9071
    Keywords: Plasminogen activator ; active oxygen ; gene expression ; radical scavengers ; endothelial cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Active oxygen, produced by cultured cells following stimulation with various growth factors, seems to be involved in signal transduction leading to cellular responses such as gene expression and growth modulation. In the present study, the intracellular oxidation state was measured in immortalized human endothelial cells (ECV304) after treatment with tumor necrosis factor (TNF)α, using a fluorescent dye and a laser-scanning confocal microscope. The intracellular oxidation state was increased 60 min after the addition of TNFα, and this increase was abolished by a radical scavenger, N-acetylcysteine (NAC), which is also a precursor of glutathione, and by pyrrolidine dithiocarbamate (PDTC). TNFα increased the steady state level of urokinase-type plasminogen activator (uPA), and NAC inhibited this increase at a dose that also inhibited the increase in the intracellular oxidation state. PDTC, on the other hand, did not affect the induction of the uPA gene by TNFα. These results suggest that intracellular glutathione level rather than the oxidation state is necessary for the induction of the uPA gene by TNFα.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01923487
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  • 4
    Electronic Resource
    Electronic Resource
    Genetic ecology: A new interdisciplinary science, fundamental for evolution, biodiversity and biosafety evaluations (1994)
    Springer
    Cellular and molecular life sciences 50 (1994), S. 429-437 
    Details
    ISSN: 1420-9071
    Keywords: Genetics ; ecology ; DNA-transfer ; conjugation ; transformation ; transduction ; transposons ; dormant cells ; epilithon ; microbial colonisation ; symbiosis ; virus resistance ; biosafety ; release of genes ; insults to humanity ; evolution ; biodiversity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Genetic ecology is the extension of our modern knowledge in molecular genetics to studies of viability, gene expression and gene movements in natural environments like soils, aquifers and digestive tracts. In such milieux, the horizontal transfer of plasmid-borne genes between phylogenetically distant species has already been found to be much more frequent than had been expected from laboratory experience. For the study of exchanges involving chromosomally-located genes, more has to be learned about the behaviour of transposons in such environments. The results expected from studies in genetic ecology are relevant for considerations of evolution, biodiversity and biosafety. The role of this new field of research in restoring popular confidence in science and in its biotechnological applications is stressed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01920741
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  • 5
    Electronic Resource
    Electronic Resource
    Expression of glycogen synthase and phosphofructokinase in muscle from Type 1 (insulin-dependent) diabetic patients before and after intensive insulin treatment (1994)
    Springer
    Diabetologia 37 (1994), S. 82-90 
    Details
    ISSN: 1432-0428
    Keywords: Type 1 (insulin-dependent) diabetes mellitus ; gene expression ; skeletal muscle ; glycogen synthase ; phosphofructokinase ; hexokinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The aim of the present study was to determine whether short-term appropriate insulinization of Type 1 (insulin-dependent) diabetic patients in longterm poor glycaemic control (HbA1C〉9.5%) was associated with an adaptive regulation of the activity and gene expression of key proteins in muscle glycogen storage and glycolysis: glycogen synthase and phosphofructokinase, respectively. In nine diabetic patients biopsies of quadriceps muscle were taken before and 24-h after intensified insulin therapy and compared to findings in eight control subjects. Subcutaneous injections of rapid acting insulin were given at 3-h intervals to improve glycaemic control in diabetic patients (fasting plasma glucose decreased from 20.8±0.8 to 8.7±0.8 mmol/l whereas fasting serum insulin increased from 59±8 to 173±3 pmol/l). Before intensified insulin therapy, analysis of muscle biopsies from diabetic patients showed a normal total glycogen synthase ctivity but a 48% decrease (p=0.006) in glycogen synthase fractional velocity (0.1 mmol/l glucose 6-phosphate) (FV0.1) and a 45% increase (p=0.01) in the half-maximal activation constant of glycogen synthase (A0.5). The activity of phosphofructokinase and the specific mRNA and immunoreactive protein levels of both glycogen synthase and phosphofructokinase were similar in the two groups. The 2.8-fold increase in serum insulin levels and the halving of the plasma glucose level for at least 15 h were associated with a normalization of glycogen synthase fractional activity (FV0.1) and of the half-maximal activation constant (A0.5) whereas the enzyme activity of phosphofructokinase and the mRNA and protein levels of both glycogen synthase and phosphofructokinase remained normal. In conclusion: 1) Reduced allosterical activation of glycogen synthase in muscle of Type 1 diabetic patients in poor metabolic control occurs in the presence of normal total activity as well as normal immunoreactive protein mass and mRNA level of glycogen synthase. 2) Changes in serum insulin within the physiological range play no role in the short-term regulation of glycogen synthase mRNA and protein abundance in muscle from Type 1 diabetic patients.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00428782
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  • 6
    Electronic Resource
    Electronic Resource
    Tissue-specific correction of lipogenic enzyme gene expression in diabetic rats given vanadate (1994)
    Springer
    Diabetologia 37 (1994), S. 1065-1072 
    Details
    ISSN: 1432-0428
    Keywords: Key words Vanadium ; lipogenic enzymes ; gene expression ; streptozotocin-diabetic rats.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Vanadium is a potent insulinomimetic agent. In vivo, its blood glucose lowering action in insulin-deficient diabetic rats is associated with corrected expression of genes involved in hepatic glucose metabolism. In this study, we investigated whether vanadate treatment also reverses the impaired expression of genes coding for key enzymes of lipogenesis in diabetic liver and white adipose tissue. Oral administration of vanadate to streptozotocin-rats caused a 55 % fall in plasma glucose levels after feeding without modifying low insulinaemia. It also partially corrected the low thyroid hormone concentrations. In untreated diabetic animals, hepatic mRNA levels of acetyl-CoA carboxylase and fatty acid synthase were reduced by more than 80 and 90 %, respectively, in close correlation with changes in enzyme activities. Three weeks of vanadate treatment totally restored acetyl-CoA carboxylase mRNA and partially restored fatty acid synthase mRNA (71 % of control levels). The activities of both lipogenic enzymes were increased 3.5 to 4-fold, to reach 45 to 65 % of control values. By contrast, in white adipose tissue, vanadate modified neither expression nor activity of both lipogenic enzymes, which remained blunted (〈 10 % of control levels). In conclusion, vanadate treatment partially restores the activities of two key lipogenic enzymes in liver, but not in white adipose tissue, of diabetic rats. This correction results from a reversal of impaired pre-translational regulatory mechanisms possibly mediated by an improvement of thyroid function and a selective restoration of liver glycolytic flux. [Diabetologia (1994) 37: 1065–1072]
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00418369
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  • 7
    Electronic Resource
    Electronic Resource
    Pancreatic Reg/pancreatic stone protein (PSP) gene expression does not correlate with beta-cell growth and regeneration in rats (1994)
    Springer
    Diabetologia 37 (1994), S. 994-999 
    Details
    ISSN: 1432-0428
    Keywords: Pancreas ; growth factors ; gene expression ; beta cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The Reg/pancreatic stone protein (PSP) gene is postulated to be an important regulator of pancreatic beta-cell growth. To investigate this hypothesis, we analysed the expression of the Reg/PSP gene following a 90% pancreatectomy and after chronic glucose infusion, two well-defined models of pancreatic beta-cell growth. There was a rapid induction of the Reg/PSP gene in the remnant pancreas after a 90% pancreatectomy in rats during the period of marked growth of the exocrine and islet tissue. However, a similar rapid, but smaller, induction of the Reg/PSP gene was observed in sham-operated rats and in non-surgical control rats in which there was no enhanced pancreatic growth. Furthermore, there was no pancreatic Reg/PSP gene induction in a model of selective beta-cell growth, the chronic glucose-infused rat. Thus, it is unlikely that Reg/PSP is a beta-cell specific growth factor, even though the function of this important pancreatic gene is still unknown.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00400462
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  • 8
    Electronic Resource
    Electronic Resource
    Pancreatic Reg/pancreatic stone protein (PSP) gene expression does not correlate with beta-cell growth and regeneration in rats (1994)
    Springer
    Diabetologia 37 (1994), S. 994-999 
    Details
    ISSN: 1432-0428
    Keywords: Key words Pancreas ; growth factors ; gene expression ; beta cells.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The Reg/pancreatic stone protein (PSP) gene is postulated to be an important regulator of pancreatic beta-cell growth. To investigate this hypothesis, we analysed the expression of the Reg/PSP gene following a 90 % pancreatectomy and after chronic glucose infusion, two well-defined models of pancreatic beta-cell growth. There was a rapid induction of the Reg/PSP gene in the remnant pancreas after a 90 % pancreatectomy in rats during the period of marked growth of the exocrine and islet tissue. However, a similar rapid, but smaller, induction of the Reg/PSP gene was observed in sham-operated rats and in non-surgical control rats in which there was no enhanced pancreatic growth. Furthermore, there was no pancreatic Reg/PSP gene induction in a model of selective beta-cell growth, the chronic glucose-infused rat. Thus, it is unlikely that Reg/PSP is a beta-cell specific growth factor, even though the function of this important pancreatic gene is still unknown. [Diabetologia (1994) 37: 994–999]
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00400462
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  • 9
    Electronic Resource
    Electronic Resource
    Tissue-specific correction of lipogenic enzyme gene expression in diabetic rats given vanadate (1994)
    Springer
    Diabetologia 37 (1994), S. 1065-1072 
    Details
    ISSN: 1432-0428
    Keywords: Vanadium ; lipogenic enzymes ; gene expression ; streptozotocin-diabetic rats
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Vanadium is a potent insulinomimetic agent. In vivo, its blood glucose lowering action in insulin-deficient diabetic rats is associated with corrected expression of genes involved in hepatic glucose metabolism. In this study, we investigated whether vanadate treatment also reverses the impaired expression of genes coding for key enzymes of lipogenesis in diabetic liver and white adipose tissue. Oral administration of vanadate to streptozotocin-rats caused a 55% fall in plasma glucose levels after feeding without modifying low insulinaemia. It also partially corrected the low thyroid hormone concentrations. In untreated diabetic animals, hepatic mRNA levels of acetyl-CoA carboxylase and fatty acid synthase were reduced by more than 80 and 90%, respectively, in close correlation with changes in enzyme activities. Three weeks of vanadate treatment totally restored acetyl-CoA carboxylase mRNA and partially restored fatty acid synthase mRNA (71% of control levels). The activities of both lipogenic enzymes were increased 3.5 to 4-fold, to reach 45 to 65% of control values. By contrast, in white adipose tissue, vanadate modified neither expression nor activity of both lipogenic enzymes, which remained blunted (〈10% of control levels). In conclusion, vanadate treatment partially restores the activities of two key lipogenic enzymes in liver, but not in white adipose tissue, of diabetic rats. This correction results from a reversal of impaired pre-translational regulatory mechanisms possibly mediated by an improvement of thyroid function and a selective restoration of liver glycolytic flux.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00418369
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  • 10
    Electronic Resource
    Electronic Resource
    Agrobacterium-mediated transformation of white mustard (Sinapis alba L.) and regeneration of transgenic plants (1994)
    Springer
    Plant cell reports 13 (1994), S. 130-134 
    Details
    ISSN: 1432-203X
    Keywords: Sinapis alba L. ; Agrobacterium tumefaciens ; transformation ; regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A procedure for the regeneration of fertile transgenic white mustard (Sinapis alba L.) is presented. The protocol is based on infection of stem explants of 7–9 day old plants with an Agrobacterium tumefaciens strain harboring a disarmed binary vector with chimeric genes encoding neomycin phosphotransferase and β-glucuronidase. Shoots are regenerated from callus-forming explants within 3–4 weeks. Under selection, 10% of the explants with transgenic embryonic callus develop into fertile transgenic plants. Rooting shoots transferred to soil yield seeds within 14–16 weeks following transformation. Integration and expression of the T-DNA encoded marker genes was confirmed by histochemical β glucuronidase assays and Southern-DNA hybridization using primary transformants and S1-progeny. The analysis showed stable integration and Mendelian inheritance of trans-genes in transformed Sinapis lines.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00239878
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  • 11
    Electronic Resource
    Electronic Resource
    Stable expression of the GUS reporter gene in chrysanthemum depends on binary plasmid T-DNA (1994)
    Springer
    Plant cell reports 14 (1994), S. 59-64 
    Details
    ISSN: 1432-203X
    Keywords: Agrobacterium ; transformation ; T-DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three chrysanthemum (Dendranthema grandiflora) cultivars were cocultivated with 2 Agrobacterium tumefaciens strains in combination with 4 pBIN19 derived binary plasmids, all carrying the Nosnptll selection gene and 35Sgus(intron) reporter gene. All binary plasmids transferred DNA to chrysanthemum explants but only pMOG410 gave good stable expression of GUS. This plasmid differs from the other plasmids in 2 aspects: 1) It carries a restored nptll gene and 2) the selection gene is positioned at the left border side of the reporter gene. Cocultivation with AGLO(pMOG410) yielded up to 13 GUS positive shoots per 100 explants. The presence of the gus and nptll gene in recovered shoots was confirmed by PCR and Southern blot analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233300
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  • 12
    Electronic Resource
    Electronic Resource
    Isolation and transformation of rice aleurone protoplasts (1994)
    Springer
    Plant cell reports 13 (1994), S. 394-396 
    Details
    ISSN: 1432-203X
    Keywords: Rice ; α-amylase ; protoplasts ; aleurone ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplasts isolated from the aleurone have been used extensively in molecular studies focusing on hormone-mediated regulation of gene expression in barley seed. To extend the use of aleurone protoplasts to other species, we have determined the conditions necessary for the isolation of protoplasts from rice aleurone layers of germinated seed. Many of the common cell wall degrading enzymes used in making protoplasts were tested for their ability to release protoplasts from rice aleurone layers. Cellulysin was found to be the most effective. Transformation of these aleurone protoplasts was accomplished using polyethylene glycol and DNA constructs containing the firefly luciferase reporter gene under the control of two different promoters were tested. Luciferase expression was 24-fold greater when the reporter gene was under the control of the CaMV 35S promoter than when the promoter from the alcohol dehydrogenase 1 gene was used. With the isolation and transformation of aleurone protoplasts from rice, it is now possible to investigate molecular events occurring in this tissue during germination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00234145
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  • 13
    Electronic Resource
    Electronic Resource
    Expression of calcium-binding protein regucalcin mRNA in rat liver is stimulated by calcitonin: The hormonal effect is mediated through calcium (1994)
    Springer
    Molecular and cellular biochemistry 136 (1994), S. 43-48 
    Details
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; rat liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The involvement of a hypocalcemic hormone calcitonin (CT) in the expression of hepatic Ca2+-binding protein regucalcin mRNA was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin complementary DNA (0.9 kb). A single oral administration of calcium chloride (100 mg Ca/100 g body weight) to rats induced a remarkable increase in the serum calcium concentration and a corresponding elevation of the liver calcium content during 120 min after the administration. Thyroparathyroidectomy (TPTX) did not cause a significant increase in the liver calcium content after calcium administration. Hepatic regucalcin mRNA level was markedly elevated by calcium administration; the level was about 180% of controls at 60 min after the administration. This increase was completely abolished by TPTX. A single subcutaneous administration of CT (synthetic eel CT; 25–100 MRC mU/100 g) to TPTX rats received oral administration of calcium (100 mg/100 g) produced a remarkable increase in hepatic regucalcin mRNA levels; the level was about 280% of controls with the dose of 25 MRC mU CT/100 g. The present finding suggests that the expression of hepatic mRNA is stimulated by CT, and that the hormonal effect is mediated through Ca2+ in rat liver.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00931603
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  • 14
    Electronic Resource
    Electronic Resource
    Expression of hepatic calcium-binding protein regucalcin mRNA is decreased by phenobarbital administration in rats (1994)
    Springer
    Molecular and cellular biochemistry 141 (1994), S. 15-19 
    Details
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; phenobarbital ; rat liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of phenobarbital on the expression of calcium-binding protein regucalcin mRNA in rat liver was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin cDNA (0.9 kb of open reading frame). Phenobarbital (4, 8 and 12 mg/ 100 g body weight) was intraperitoneally administered to rats 3 times with 24 h intervals, and the animals were sacrificed by bleeding at 24 h after the last administration. The hepatic regucalcin mRNA levels were markedly reduced by phenobarbital administration. This decrease was about 50% of control level with the 12 mg/100 g dose. Moreover, the hepatic regucalcin concentration was significantly decreased by the administration of phenobarbital (12 mg/100 g), although the serum regucalcin concentration was not altered appreciably. Meanwhile, serum transaminases (GOT and GPT) activities were not increased by the administration of phenobarbital (4 and 12 mg/100 g). The present study demonstrates that the expression of hepatic regucalcin mRNA is decreased by phenobarbital administration in rats, suggesting that regucalcin does not have a role in drug metabolism related to phenobarbital.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00935586
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  • 15
    Electronic Resource
    Electronic Resource
    Regulation of mRNA transcripts and DNA synthesis in the rat heart following intravenous injection of transforming growth factor beta1 (1994)
    Springer
    Molecular and cellular biochemistry 141 (1994), S. 145-151 
    Details
    ISSN: 1573-4919
    Keywords: pressure overload ; myocardium ; gene expression ; fibroblast ; extracellular matrix ; ventricular hypertrophy ; growth factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Transforming Growth Factor-beta1 (TGF-β1) is expressed in the heart by muscle and non-muscle cardiac cells.In vitro, cardiac myocytes and non-muscle cells including cardiac fibroblasts and endothelial cells respond to regulatory effects of TGF-β1. Expression of TGF-β1 in the heart is subject to regulation by hemodynamic stimuli. Increased expression of mRNA transcripts for TGF-β1 has been reported in several models of cardiac hypertrophy. The objective of this study was to determine the effect of TGF-β1 in the myocardium. TGF-β1 was injected intravenously. Expression of mRNA transcripts for functional and structural proteins was determined by Northern hybridization analysis. DNA synthesis was determined by measurement of3H-thymidine incorporation into ventricular DNA. The results showed differential regulation of mRNAs for myocyte- and non-myocyte-specific proteins in the heart of TGF-β1 treated rats. Moderate but statistically significant decrease in DNA synthesis was observed in the heart of TGF-β1 treated rats (37.5%, P〈0.025). Together, these data point to a physiological role for TGF-β1 in the heart. They further suggest that similar to its diversein vitro cell-specific regulatory effects, TGF-β1 may have multicellular targets in the heart. Effect of TGF-β1 alone or combined with those of other cytokines/hormones that come into play, as the result of its administration, may be responsible for altered gene expression and DNA synthesis in the myocardium. We propose that in experimental models of myocardial hypertrophy which are associated with increased expression of TGF-β1 in the heart, the contribution of regulatory effects of this growth factor to the manifestations of ventricular hypertrophy could be significant.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00926178
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  • 16
    Electronic Resource
    Electronic Resource
    Calcium and calcium-binding proteins in the nucleus (1994)
    Springer
    Molecular and cellular biochemistry 135 (1994), S. 79-88 
    Details
    ISSN: 1573-4919
    Keywords: calcium ; nucleus ; calpain ; calmodulin ; cell division ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Calcium has long been known to play a role as a key cytoplasmic second messenger, but until relatively recently its possible involvement in nuclear signal transduction and the regulation of nuclear events has not been extensively studied. Evidence revealing the presence of transmembrane nuclear Ca2+ gradients and a variety of intranuclear Ca2+ binding proteins has fueled renewed interest in this key ion and its involvement in cell-cycle timing and division, gene expression, and protein activation. This review will offer an overview of the current state of knowledge and theory regarding calcium orchestration of nuclear functions and events and discuss possible future directions in this field of study.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00925963
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  • 17
    Electronic Resource
    Electronic Resource
    Transcriptional regulation and autoregulation of the human gene for ADP-ribosyltransferase (1994)
    Springer
    Molecular and cellular biochemistry 138 (1994), S. 99-104 
    Details
    ISSN: 1573-4919
    Keywords: poly(ADP-ribosyl) transferase (human) ; autoregulation ; gene expression ; promoter structure ; cruciform structure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Human nuclear poly(ADP-ribosyl)transferase (ADPRT) modifies proteins with branched ADP-ribose-polymers. Various proteins, including ADPRT itself, serve as acceptors for polyADP-ribose. Target proteins include those controlling basic cellular processes such as DNA repair, differentiation and proliferation. Because of the outstanding features of this enzyme: automodification, several functional domains and central role in physiology of the cell, the molecular biology of ADPRT gained wide interest. The promoter structure contains several CCAAT/TATA boxes and SP1 sites. However, there is no CCAAT/TATA box in the neighbourhood of an SP1 site and, thus no obvious site for initiation of transcription. Within this region there are several noteworthy inverted repeats, which by internal basepairing could form two types of cruciform structures. Deletion analysis revealed that these cruciform structures have functional significance. Removal of one type increases the promoter activity, whereas removal of the other diminishes the promoter function. Overexpression of ADPRT from heterologous promoters (MMTV, SV40) leads to repression of the activity of the ADPRT promoter. Indeed, ADPRT was shown to bind specifically to one type of cruciform structure. This specific interaction indicates autorepression of the ADPRT gene: the enzyme ADPRT acts directly as a negative modulator of the activity of its own promoter.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00928449
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  • 18
    Electronic Resource
    Electronic Resource
    Expression of the mitochondrial creatine kinase genes (1994)
    Springer
    Molecular and cellular biochemistry 133-134 (1994), S. 235-243 
    Details
    ISSN: 1573-4919
    Keywords: creatine kinase ; mitochondria ; metabolism ; creatine phosphate shuttle ; gene expression ; muscle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Mitochondrial Creatine Kinase (MtCK) is responsible for the transfer of high energy phosphate from mitochondria to the cytosolic carrier, creatine, and exists in mammals as two isoenzymes encoded by separate genes. In rats and humans, sarcomere-specific MtCK (sMtCK) is expressed only in skeletal and heart muscle, and has 87% nucleotide identity across the 1257 bp coding region. The ubiquitous isoenzyme of MtCK (uMtCK) is expressed in many tissues with highest levels in brain, gut, and kidney, and has 92% nucleotide identity between the 1254 bp coding regions of rat and human. Both genes are highly regulated developmentally in a tissue-specific manner. There is virtually no expression of sMtCK mRNA prior to birth. Unlike cytosolic muscle CK (MCK) and brain CK (BCK), there is no developmental isoenzyme switch between the MtCKs. Cell culture models representing the tissue-specific expression of either sMtCK or uMtCK are available, but there are no adequate developmental models to examine their regulation. Several animal models are available to examine the coordinate regulation of the CK gene family and include 1) Cardiac Stress by coarctation (sMtCK, BCK, and MCK), 2) Uterus and placenta during pregnancy (uMtCK and BCK), and 3) Diabetes and mitochondrial myopathy (sMtCK, BCK, and MCK). We report the details of these findings, and discuss the coordinate regulation of the genes necessary for high-energy transduction.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01267957
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  • 19
    Electronic Resource
    Electronic Resource
    Nuclear Ca2+: physiological regulation and role in apoptosis (1994)
    Springer
    Molecular and cellular biochemistry 135 (1994), S. 89-98 
    Details
    ISSN: 1573-4919
    Keywords: calcium ; cell death ; nuclei ; apoptosis ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The last decade has seen the rapid development of research investigating the molecular mechanisms whereby hormones, peptide growth factors and cytokines regulate cell metabolism, differentiation and proliferation. One general signalling mechanism used to transfer the information delivered by agonists into appropriate intracellular compartments involves the rapid Ca2+ redistribution throughout the cell, which results in transient elevations of the cytosolic free Ca2+ concentration. Ca2+ signals are required for a number of cellular processes including the activation of nuclear processes such as gene transcription and cell cycle events. The latter require that appropriate Ca2+ signals elicited in response to agonists be transduced across the nuclear envelope. It has generally been assumed that small molecules, metabolites and ions could freely diffuse across the nuclear envelope. Nevertheless several findings during the past few years have suggested that nuclear pore permeability can be regulated and that ion transport systems and ion-selective channels may exist on the nuclear membranes and regulate intranuclear processes. Intranuclear Ca2+ fluctuations can affect chromatin organization, induce gene expression and also activate cleavage of nuclear DNA by nucleases during programmed cell death or apoptosis. The possible mechanisms involved in nuclear Ca2+ transport and the control of nuclear Ca2+-dependent enzymes in apoptosis is discussed below.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00925964
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  • 20
    Electronic Resource
    Electronic Resource
    A retinoic acid-inducible skin-specific gene (RIS-1/psoriasin): molecular cloning and analysis of gene expression in human skinin vivo and cultured skin cellsin vitro (1994)
    Springer
    Molecular biology reports 20 (1994), S. 75-83 
    Details
    ISSN: 1573-4978
    Keywords: retinoic acid ; skin ; differential hybridization ; cloning ; keratinocytes ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A retinoic acid (RA) inducible skin-specific gene transcript (RIS-1) was isolated by differential hybridization screening of a RA-treated human skin cDNA library. The library was constructed from pooled RNA derived from normal adult human skin treated with alltrans-RA for 4 h (n=6) and 12 h (n=6)in vivo. RIS-1 cDNA corresponded to a 0.6 kb transcript that was barely detectable in normal adult human skin but was significantly induced by 8 h in RA-treated compared to vehicle-treated skin (range 1.1–3.6 fold). Prolonged RA treatment for up to 24 h further increased relative RIS-1 mRNA levels by 1.3–5.5 fold. HPLC analysis of the RA content of 0.1% RA-treated skinin vivo revealed significant levels at 6 h (18.8–120.6 ng RA/g wet weight tissue; approximately 240 nM), immediately preceding the time point at which the increased RIS-1 mRNA level was first seen. This concentration of RA also induced the mRNA levels for cellular RA binding protein II (1.6–19 fold), a marker of RA activity in human skin. RIS-1 mRNA was detected by Northern and dot blotting only in normal skin but not in any other normal human tissues examined, indicating a tissue-specific pattern of gene expression. RIS-1 transcripts were detected at very low levels in untreated cultured human epidermal keratinocytes, while no expression was seen in dermal fibroblasts and melanocytes, the other major cell types in skin. Southern analysis of human and mouse DNA indicated the existence of evolutionarily conserved sequences for RIS-1 between these two species. The polypeptide sequence derived from the partial RIS-1 cDNA was found to be identical to the calcium binding domain found in ‘psoriasin’, a gene whose expression appears to be increased in the skin of psoriasis patients.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00996356
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  • 21
    Electronic Resource
    Electronic Resource
    Expression of nitrate assimilation related genes in Chlamydomonas reinhardtii (1994)
    Springer
    Plant molecular biology 24 (1994), S. 185-194 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; light/nitrate regulation ; nitrate reductase ; nitrate transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The mRNA accumulation pattern of the Chlamydomonas reinhardtii nitrate assimilation-related gene cluster has been elucidated. In ammonium-grown wild-type cells, nit-1 (nitrate reductase, NR), nar-1, nar-2 and nar-3 (nitrate transporter) genes showed very similar kinetics of expression when transferred to nitrate medium. Transcripts of all these genes accumulated transiently in ammonium-grown wild-type cells after a one-hour incubation in nitrogen-free medium, and practically disappeared at about 2 hours. Mutant strains lacking functional nitrate reductase showed similar accumulation kinetics of these transcripts during both nitrate induction and derepression in nitrogen-free media. In contrast to the other nar transcripts, that nar-4, a gene sharing similar sequences with nar-3, accumulated in small amounts in wild-type cells, and only increased after a long nitrate induction period. Nitrate and light showed a strong positive effect on the accumulation of nit-1 gene transcripts. Acetate as a carbon source allowed accumulation of nit-1 mRNA in the dark, indicating the existence of interactions between light and carbon metabolism in nit-1 gene expression. Our data strongly suggest that NR negatively autoregulates its own expression and that of nar genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040584
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  • 22
    Electronic Resource
    Electronic Resource
    Expression of one of the members of the Arabidopsis chaperonin 60β gene family is developmentally regulated and wound-repressible (1994)
    Springer
    Plant molecular biology 24 (1994), S. 195-202 
    Details
    ISSN: 1573-5028
    Keywords: plant transformation ; chaperonin 60β ; β-glucuronidase ; wound repression ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To study the pattern of gene regulation of the plastid chaperonin 60β gene family a chimaeric gene was constructed fusing the 5′-flanking region of the chaperonin 60β B3 gene to the β-glucuronidase reporter gene. Histochemical and fluorometric analysis of the GUS activity present in transgenic plants harbouring this gene construct showed that the B3 promoter is expressed in leaves, stem, petioles and several flower tissues. The pattern of cell type-specific expression in stems and flowers was found to be developmentally regulated. Expression of the B3 promoter was found not to be heat-inducible, but highly repressed by wounding. The rapid decay in GUS activity upon wounding indicates that, at least under some physiological conditions, the gene product of this reporter gene is not as stable as has been previously thought.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040585
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  • 23
    Electronic Resource
    Electronic Resource
    Low-temperature-responsive barley genes have different control mechanisms (1994)
    Springer
    Plant molecular biology 24 (1994), S. 879-888 
    Details
    ISSN: 1573-5028
    Keywords: barley ; cold acclimation ; gene expression ; low temperature genes ; nuclear run-on transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Several low-temperature-responsive (LTR) genes from barley have been shown to have high steady-state transcript levels. Run-on transcription was used to determine the control of expression of these LTR genes. Six of these are shown to be transcriptionally regulated (blt 4/9, blt 101, blt 1015, blt 63, blt 49, blt 410) whilst three are post-transcriptionally regulated (blt 14, blt 411, blt 801). Two transcriptionally regulated genes (blt 4/9 and blt 101) and one post-transcriptionally regulated gene (blt 14) have been used in expression studies. The time course for the appearance and decay of these transcripts is given. Initial appearance and steady-state levels of individual transcripts have different temperature characteristics but no single gene correlates with the cold acclimation response. We suggest that these different response profiles may represent a means of fine-tuning the low-temperature response. One gene, blt 4/9, also accumulated high steady-state levels of transcript in response to drought and a nutrient stress. However, only drought has an acclimating effect on barley plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014442
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  • 24
    Electronic Resource
    Electronic Resource
    The 24kDa outer envelope membrane protein from spinach chloroplasts: molecular cloning, in vivo expression and import pathway of a protein with unusual properties (1994)
    Springer
    Plant molecular biology 25 (1994), S. 167-177 
    Details
    ISSN: 1573-5028
    Keywords: Spinacia oleracea ; chemical cleavage ; gene expression ; polymerase chain reaction ; protein transport ; SDS-PAGE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The 24 kDa outer envelope membrane protein of spinach chloroplasts (omp24) represents a major constituent of this membrane. Sequences of tryptic and endoprotease Glu-C peptides derived from omp24 allowed the design of oligonucleotides which were used to generate a DNA fragment by polymerase chain reaction using spinach cDNA as template. This fragment served as a probe to screen a cDNA library for a full-length clone of the omp24 coding sequence. The protein predicted from the complete sequence only has 148 amino acids and a molecular mass of 16294 Da. It is an acidic protein (calculated isoelectric point 4.8) with a high content of proline residues. Expression of the coding sequence in Escherichia coli and characterization of the purified recombinant protein produced revealed that the overestimation of its molecular mass by SDS-PAGE (ca. 25 kDa) is due to its abnormal amino acid composition. Despite its rather low hydrophobicity (polarity index 49%), omp24 appears to be deeply embedded in the outer membrane. Insertion of omp24 into the membrane proceeds almost independently of surface receptors or targeting sequence but, in contrast to other known outer envelope membrane proteins, is stimulated by ATP.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023235
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  • 25
    Electronic Resource
    Electronic Resource
    Gene expression of nitrite reductase in Scots pine (Pinus sylvestris L.) as affected by light and nitrate (1994)
    Springer
    Plant molecular biology 25 (1994), S. 449-457 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; light ; nitrate ; nitrite reductase ; Pimus sylvestris L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A partial cDNA clone (PSnir) encoding the C-terminal region of nitrite reductase was isolated from a λgt 11 library of the gymnospermPimus sylvestris (L.). Nucleotide sequence analysis showed that PSnir contains a reading frame encoding 105 amino acid residues. The amino acid sequence revealed a homology to NiR of 63–68% to dicotyledoneous and of 57–59% to monocotyledoneous species. The protein region implicated to be involved in binding of the prosthetic group is highly conserved between the NiR of the gymnosperm and of angiosperms. In all organs (cotyledonary whorls, hypocotyls, roots) the pattern of NiR gene expression in response to nitrate and light is the same at the level of transcript accumulation and at the enzyme level. This suggests that regulation of NiR gene expression in the Scots pine seedling is predominantly at the level of transcript accumulation. The highest NiR appearance was observed in roots and hypocotyls. In the cotyledonary whorls only small amounts of NiR were found. In roots and hypocotyls the accumulation of NiR mRNA and the appearance of NiR protein is mainly controlled by nitrate, whereas the regulation of NiR gene expression in the whorls is strongly affected by light and the inducive effect of nitrate is only weak.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00043873
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  • 26
    Electronic Resource
    Electronic Resource
    Isolation and characterization of two tightly linked catalase genes from castor bean that are differentially regulated (1994)
    Springer
    Plant molecular biology 25 (1994), S. 507-516 
    Details
    ISSN: 1573-5028
    Keywords: castor bean (Ricinus communis) ; catalase gene ; gene expression ; germination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two catalase genes,cat1 andcat2, have been isolated from the castor bean genome. They were located in the same direction on a chromosome at a distance of 2.4 kb,cat1 being on the downstream side ofcat2. The two genes contained introns at the same positions except that one of the 7 introns incat1 is missing incat2 and the corresponding introns differed in size and sequence between the two genes. The translated regions of the two genes had the same number of nucleotides and exhibited 81.3% nucleotide sequence identity. In addition to introns, the nucleotide sequences of the 5′-and 3′-flanking regions are highly divergent between the two genes. In etiolated seedlings,cat1 mRNA was present abundantly in endosperms and cotyledons and only in a small amount in roots. Thecat1 mRNA could not be detected in hypocotyls. By contrast,cat2 mRNA is most abundant in hypocotyls and roots, while endosperms and cotyledons contained only low levels ofcat2 mRNA. Although neithercat1 norcat2 mRNA could be detected in dry seeds, both mRNAs showed temporal accumulation in the endosperm in response to germination. These results suggest that expression of two tightly linked catalase genes of castor bean,cat1 andcat2, are differentially regulated during development.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00043878
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  • 27
    Electronic Resource
    Electronic Resource
    Molecular improvement of cereals (1994)
    Springer
    Plant molecular biology 25 (1994), S. 925-937 
    Details
    ISSN: 1573-5028
    Keywords: cereals ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014667
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  • 28
    Electronic Resource
    Electronic Resource
    Characterization of four new β-tubulin genes and their expression during male flower development in maize (Zea mays L.) (1994)
    Springer
    Plant molecular biology 24 (1994), S. 295-315 
    Details
    ISSN: 1573-5028
    Keywords: β-tubulin ; microtubules ; maize ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four different β-tubulin coding sequences were isolated from a cDNA library prepared from RNA from maize seedling shoots. The four genes (designated tub4, tub6, tub7 and tub8) represented by these cDNA clones together with the tub1 and tub2 genes reported previously encode six β-tubulin isotypes with 90–97.5% amino acid sequence identity. Results from phylogenetic analysis of 17 β-tubulin genes from monocot and dicot plant species indicated that multiple extant lines of β-tubulin genes diverged from a single precursor after the appearance of the two major subfamilies of α-tubulin genes described previously. Hybridization probes from the 3′ non-coding regions of six β-tubulin clones were used to quantify the levels of corresponding tubulin transcripts in different maize tissues including developing anthers and pollen. The results from these dot blot hybridization experiments showed that all of the β-tubulin genes were expressed in most tissues examined, although each gene showed a unique pattern of differential transcript accumulation. The tub1 gene showed a high level of transcript accumulation in meristematic tissues and almost no accumulation in the late stages of anther development and in pollen. In contrast, the level of tub4 transcripts was very low during early stages of male flower development but increased markedly (more than 100 times) during the development of anthers and in pollen. Results from RNAse protection assays showed that this increased hybridization signal resulted from expression of transcripts from one or two genes closely related to tub4. The tub4-related transcripts were not present in shoot tissue. Transcripts from the tub2 gene accumulated to very low levels in all tissues examined, but reached the highest levels in young anthers containing microspore mother cells. RNAse protection assays were used to measure the absolute levels of α- and β-tubulin transcripts in seedling shoot and in pollen. The α-tubulin gene subfamily I genes (tua1, tua2, tua4) contributed the great majority of α-tubulin transcripts in both shoot and pollen. Transcripts from the β-tubulin genes tub4, tub6, tub7, and tub8 were predominant in shoot, but were much less significant than the tub4-related transcripts in pollen.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00020169
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  • 29
    Electronic Resource
    Electronic Resource
    Structural and functional analysis of an Opaque-2-related gene from sorghum (1994)
    Springer
    Plant molecular biology 24 (1994), S. 515-523 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; b-ZIP motif ; seed storage proteins ; trans-acting factors ; transcription factors ; transcriptional regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Opaque-2 (O2) gene from maize encodes a transcriptional activator of the b-ZIP class. We have isolated and characterized a gene from sorghum, related in sequence to the O2 gene from maize. A single copy of the gene is present in sorghum. Both genomic and cDNA sequences of the O2-related sorghum gene were determined. The sequence is highly homologous to maize O2 both in the promoter and in the coding region. The most closely related sequences contain the b-ZIP domain with only 11 amino acid substitutions in a total of 122 residues. In transient expression assays, the sorghum O2-related coding sequence, expressed from a CaMV 35S promoter, activates expression from the maize b-32 promoter as effectively as that obtained with the maize O2 sequence.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00024119
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  • 30
    Electronic Resource
    Electronic Resource
    Cloning of two gibberellin-regulated cDNAs from Arabidopsis thaliana by subtractive hybridization: expression of the tonoplast water channel, γ-TIP, is increased by GA3 (1994)
    Springer
    Plant molecular biology 24 (1994), S. 603-615 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; dwarf mutant ; gene expression ; gibberellin ; subtractive hybridization ; tonoplast intrinsic protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Arabidopsis ga1 mutant has very low levels of endogenous, active gibberellins and thus has an extreme dwarf phenotype; application of GA3 induces stem elongation and flower development. To test the hypothesis that GA action in this system involves changes in gene expression, we have cloned mRNAs whose abundance changes following GA application. A subtraction cloning scheme for the isolation of differentially regulated cDNAs was established, involving hybridization of single-stranded cDNA to biotinylated mRNA. cDNA populations enriched up to 150-fold in GA-regulated sequences were produced and cDNA libraries generated. Screening of these libraries has isolated two clones that identify mRNAs of ca. 1100 and 750 bases whose abundance is markedly increased 24 h after GA application. One of these clones encodes the vegetative form of the Arabidopsis tonoplast intrinsic protein (γ-TIP), a water channel protein, the expression of which has recently been shown to be correlated with regions of cell expansion. The second clone is expressed only in the inflorescence and encodes a proline- and glycine-rich protein that may be a cell wall component.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023557
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  • 31
    Electronic Resource
    Electronic Resource
    Identification of conserved domains in the Δ12 desaturases of cyanobacteria (1994)
    Springer
    Plant molecular biology 24 (1994), S. 643-650 
    Details
    ISSN: 1573-5028
    Keywords: Anabaena variabilis ; fatty-acid desaturation ; Synechococcus PCC7002 ; Synechococcus PCC7942 ; Synechocystis PCC6714 ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cyanobacterial genes for enzymes that desaturate fatty acids at the Δ12 position, designated desA, were isolated from Synechocystis PCC6714, Synechococcus PCC7002 and Anabaena variabilis by crosshybridization with a DNA probe derived from the desA gene of Synechocystis PCC6803. The genes of Synechocystis PCC6714, Synechococcus PCC7002 and A. variabilis encode proteins of 349, 347 and 350 amino acid residues, respectively. The transformation of Synechococcus PCC7942 with the desA genes from Synechocystis PCC6714, Synechococcus PCC7002 and A. variabilis was associated with the ability to introduce a second double bond at the Δ12 position of fatty acids. The amino acid sequence of the products of the desA genes revealed the presence of four conserved domains. Since one of the conserved domains was also found in the amino acid sequences of ω3 desaturases of Brassica napus and mung bean, this domain may play an essential role in the introduction of a double bond into fatty acids bound to membrane lipids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023560
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  • 32
    Electronic Resource
    Electronic Resource
    Directed disruption of the Chlamydomonas chloroplast psbK gene destabilizes the photosystem II reaction center complex (1994)
    Springer
    Plant molecular biology 24 (1994), S. 779-788 
    Details
    ISSN: 1573-5028
    Keywords: Chlamydomonas reinhardtii ; chloroplast ; transformation ; photosystem II ; psbK
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using particle gun-mediated chloroplast transformation we have disrupted the psbK gene of Chlamydomonas reihardtii with an aadA expression cassette that confers resistance to spectinomycin. The transformants are unable to grow photoautotrophically, but they grow normally in acetate-containing medium. They are deficient in photosystem II activity as measured by fluorescence transients and O2 evolution and they accumulate less than 10% of wild-type levels of photosystem II as measured by immunochemical means. Pulse-labeling experiments indicate that the photosystem II complex is synthesized normally in the transformants. These results differ from those obtained previously with similar cyanobacterial psbK mutants that were still capable of photoautotrophic growth (Ikeuchi et al., J. Biol. Chem. 266 (1991) 1111–1115). In C. reinhardtii the psbK product is required for the stable assembly and/or stability of the photosystem II complex and essential for photoautotrophic growth. The data also suggest that the stability requirements of the photosynthetic complexes differ considerably between C. reinhardtii and cyanobacteria.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00029859
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  • 33
    Electronic Resource
    Electronic Resource
    A phenylalanine ammonia-lyase gene from melon fruit: cDNA cloning, sequence and expression in response to development and wounding (1994)
    Springer
    Plant molecular biology 26 (1994), S. 473-479 
    Details
    ISSN: 1573-5028
    Keywords: Cucumis melo ; melon ; phenylalanine ammonia-lyase ; gene expression ; ripening ; wounding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phenylalanine ammonia-lyase (PAL) is the first enzyme of phenylpropanoid biosynthesis involved in the synthesis of a multiplicity of plant natural products. We have isolated and characterized a nearly fulllength cDNA clone (pmPAL-1) corresponding to a melon fruit (Cucumis melo L. var. reticulatus) gene coding for a protein which is highly similar to PAL from other lants. Melon fruit PAL is transcriptionally induced both in response to fruit ripening and wounding. PAL gene expression follows the kinetics of expression of the ethylene biosynthetic genes during fruit development. In contrast, ethylene biosynthetic genes show different induction kinetics compared to PAL expression in response to wounding. Similar results have been found for two other genes coding for enzymes involved in flavonoid biosynthesis (chalcone synthase, CHS; chalcone isomerase, CHI). Our results imply that regulation of defense gene expression in melon is a co-ordinated process in response to both ethylene and an ethylene-independent wound signal.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00039557
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  • 34
    Electronic Resource
    Electronic Resource
    Regulation of an Arabidopsis oleosin gene promoter in transgenic Brassica napus (1994)
    Springer
    Plant molecular biology 25 (1994), S. 193-205 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis ; embryo ; gene expression ; oleosin ; promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Progressive deletions of the 5′-flanking sequences of an Arabidopsis oleosin gene were fused to β-glucuronidase (GUS) and introduced into Brassica napus plants using Agrobacterium-mediated transformation. The effect of these deletions on the quantitative level of gene expression, organ specificity and developmental regulation was assessed. In addition, the influence of abscisic acid (ABA), jasmonic acid (JA), sorbitol and a combined ABA/sorbitol treatment on gene expression was investigated. Sequences that positively regulate quantitative levels of gene expression are present between −1100 to −600 and −400 to −200 of the promoter. In addition, sequences present between −600 and −400 down-regulate quantitative levels of expression. In transgenic B. napus plants, the oleosin promoter directs seed-specific expression of GUS which is present at early stages of seed development and increases throughout seed maturation. Sequences present between −2500 and −1100 of the promoter are involved in modulating the levels of expression at early stages of embryo development. Histochemical staining of embryos demonstrated that expression is uniform throughout the tissues of the embryo. Sequences involved in the response to ABA and sorbitol are present between −400 and −200. The induction of GUS activity by a combined ABA/sorbitol treatment is additive suggesting that ABA is not the sole mediator of osmotically induced oleosin gene expression. A response to JA was only observed when the oleosin promoter was truncated to −600 suggesting that the reported effect of JA on oleosin gene expression may be at a post-transcriptional level.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023237
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  • 35
    Electronic Resource
    Electronic Resource
    Characterization of a cDNA from Arabidopsis thaliana encoding a potential thiol protease whose expression is induced independently by wilting and abscisic acid (1994)
    Springer
    Plant molecular biology 25 (1994), S. 259-270 
    Details
    ISSN: 1573-5028
    Keywords: abscisic acid ; Arabidopsis thaliana ; gene expression ; mutants ; signal transduction ; stress ; thiol protease ; wilting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The sequence and expression characteristics are described of a wilt-inducible gene in Arabidopsis thaliana. A 1494 encodes a potential thiol protease whose mRNA accumulates rapidly in shoot tissue upon the loss of turgor. A1494 mRNA levels peaked after ca. 4 h and declined thereafter. Dehydration also induced rapid biosynthesis of the phytohormone abscisic acid (ABA), which continued for at least 9 h. Exogenous ABA induced the accumulation of A1494 mRNA, with kinetics similar to those after wilting. Rehydration of wilted shoots led to a rapid decline in the content of both ABA and A1494 mRNA. Wilting and ABA independently induced A1494 expression as evidenced by the effects of ABA and wilting on the ABA-deficient aba-1 and ABA-insensitive abi-1 and abi-3 genotypes. A1494 mRNA was not detectable in aba-1 shoots but accumulated rapidly after either wilting or ABA treatment, whereas the shoot ABA content was increased only by ABA treatment. ABA had no effect on A1494 mRNA levels in the abi-1 and abi-3 mutants but wilting did result in enhanced A1494 expression. Heat shock had only a minor effect on A1494 mRNA levels, whereas exposure to low temperature resulted in substantial accumulation of A1494 mRNA in wild-type shoots. However, this latter response, unlike that to drought, was mediated exclusively via ABA synthesis as demonstrated by the lack of A1494 mRNA accumulation in cold-treated aba-1 shoots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023242
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  • 36
    Electronic Resource
    Electronic Resource
    Stable transformation and long-term expression of the gusA reporter gene in callus lines of perennial ryegrass (Lolium perenne L.) (1994)
    Springer
    Plant molecular biology 24 (1994), S. 401-405 
    Details
    ISSN: 1573-5028
    Keywords: Lolium perenne L. ; transformation ; rice gene GOS2 ; long-term GUS expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Stable transformation of perennial ryegrass (Lolium perenne L.) was achieved by biolistic bombardment of a non embryogenic cell suspension culture, using the hpt and gusA gene. The transformation yielded on the average 5 callus lines per bombardment (1.4×106 cells). Stable integration of the genes into the plant genome was demonstrated by Southern analysis of DNA, isolated from hygromycin-resistant callus lines. The gusA reporter gene, which was regulated by the constitutive promoter of the rice gene GOS2, was expressed in both transient and stable transformation assays, indicating that this promoter is suitable for expression of a transferred gene in perennial ryegrass. Long-term GUS expression was observed in ca. 40% of the callus lines, whereas the other callus lines showed instability after 6 months and 1 year of culture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00020178
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  • 37
    Electronic Resource
    Electronic Resource
    Tissue-specific expression of the large ATP synthase gene cluster in spinach plastids (1994)
    Springer
    Plant molecular biology 25 (1994), S. 369-376 
    Details
    ISSN: 1573-5028
    Keywords: ATP synthase ; chloroplast ; gene expression ; plastid ; RNA stability ; transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plastids present in different tissues may vary morphologically and functionally, despite the fact that all plastids within the same plant contain identical genomes. This is achieved by regulation of expression of the plastid genome by tissue-specific factors, the mechanisms of which are not fully understood. The proton translocating ATP synthase/ATPase is a multisubunit complex composed of nine subunits, six encoded in the plastid and three in the nucleus. We have investigated the tissue-specific expression of the large ATP synthase gene cluster in spinach (Spinacia oleracea). This gene cluster encodes four of the six plastid-encoded ATP synthase genes. Transcript abundance, transcriptional activity, and transcript stability were investigated relative to gene dosage in root plastids and in stem, leaf, and flower chloroplasts. All three of these factors display significant tissue-specific variation. It was intriguing to discover that, although transcript abundance normalized to gene dosage varies in each tissue, transcript abundance as a proportion of the entire plastid RNA population in each tissue is not significantly different. Thus it appears that in these tissues the variation in transcription and stability of transcripts derived from the large ATP synthase gene cluster balances to yield an equivalent proportion of these transcripts in the plastid RNA population. Expression of this gene cluster in photosynthetic as well as non-photosynthetic tissues may facilitate the plasticity of structure and function which is characteristic of plastids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00043866
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  • 38
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    Electronic Resource
    Structure and function of selectable and non-selectable transgenes in maize after introduction by particle bombardment (1994)
    Springer
    Plant molecular biology 25 (1994), S. 951-961 
    Details
    ISSN: 1573-5028
    Keywords: Cell biology ; epigenetics ; maize ; transformation ; transgenes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Zea mays transformants produced by particle bombardment of embryogenic suspension culture cells of the genotype A188 × B73 and selected on kanamycin or bialaphos were characterized with respect to transgene integration, expression, and inheritance. Selection on bialaphos, mediated by thebar orpat genes, was more efficient than selection on kanamycin, mediated by thenptII gene. Most transformants contained multicopy, single locus, transgene insertion events. A transgene expression cassette was more likely to be rearranged if expression of that gene was not selected for during callus growth. Not all plants regenerated from calli representing single transformation events expressed the transgenes, and a non-selectable gene (uidA) was expressed in fewer plants than was the selectable transgene. Mendelian inheritance of transgenes consistent with transgene insertion at a single locus was observed for approximately two thirds of the transformants assessed. Transgene expression was typically, but not always, predictable in progeny plants-transgene silencing, as well as poor transgene transmission to progeny, was observed in some plant lines in which the parent plants had expressed the transgene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014669
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  • 39
    Electronic Resource
    Electronic Resource
    The Chlorella virus adenine methyltransferase gene promoter is a strong promoter in plants (1994)
    Springer
    Plant molecular biology 26 (1994), S. 85-93 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; monocot cells ; promoter strength ; transient expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An upstream region isolated from a eukaryotic algal virus adenine methyltransferase gene was tested for promoter function in plants. Fusion of this region to the chloramphenicol acetyltransferase reporter gene resulted in significantly higher expression than fusion with the cauliflower mosaic virus 35S promoter. Strong levels of expression were also found in electroporated monocot plant cells. The promoter activity in transgenic tobacco plants showed tissue-specific expression. Leaves had the highest expression followed by stems and flowers. The promoter activity was not detected in root tissue. Environmental cues, such as light, and the phytohormones auxin and cytokinines had no effect on the promoter's expression. This promoter might be utilized to achieve high levels of expression of introduced genes in higher plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00039522
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  • 40
    Electronic Resource
    Electronic Resource
    Cloning of ω3 desaturase from cyanobacteria and its use in altering the degree of membrane-lipid unsaturation (1994)
    Springer
    Plant molecular biology 26 (1994), S. 249-263 
    Details
    ISSN: 1573-5028
    Keywords: desB gene ; desaturase ; fatty acid ; Synechococcus sp. PCC 7942 ; Synechocystis sp. PCC 6803 ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cyanobacteria respond to a decrease in temperature by desaturating fatty acids of membrane lipids to compensate for the decrease in membrane fluidity. Among various desaturation reactions in cyanobacteria, the desaturation of the ω3 position of fatty acids is the most sensitive to the change in temperature. In the present study, we isolated a gene, designated desB, for the ω3 desaturase from the cyanobacterium, Synechocystis sp. PCC 6803. The desB gene encodes a protein a 359 amino-acid residues with molecular mass of 41.9 kDa. The desB gene is transcribed as a monocistronic operon that produced a single transcript of 1.4 kb. The level of the desB transcript in cells grown at 22°C was 10 times higher than that in cells grown at 34°C. In order to manipulate the fatty-acid unsaturation of membrane lipids, the desB gene in Synechocystis sp. PCC 6803 was mutated by insertion of a kanamycin-resistance gene cartridge. The resultant mutant was unable to desaturate fatty acids at the ω3 position. The desA gene, which encodes the Δ12 desaturase of Synechocystis sp. PCC 6803, and the desB gene were introduced into Synechococcus sp. PCC 7942. Whilst the parent cyanobacterium can only desaturate membrane lipids at the Δ9 position of fatty acids, the resultant transformant was able to desaturate fatty acids of membrane lipids at the Δ9, Δ12 and ω3 positions. These results confirm the function of the desB gene and demonstrate that it is possible to genetically manipulate the fatty-acid unsaturation of membrane lipids in cyanobacteria.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00039536
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  • 41
    Electronic Resource
    Electronic Resource
    Meristem-specific gene expression directed by the promoter of the S-phase-specific gene, cyc07, in transgenic Arabidopsis (1994)
    Springer
    Plant molecular biology 24 (1994), S. 863-878 
    Details
    ISSN: 1573-5028
    Keywords: cell cycle ; gene expression ; meristem ; promoter analysis ; transgenic Arabidopsis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A genomic clone for the cyc07 gene, which is expressed specifically at the S phase during the cell cycle in synchronous cultures of periwinkle (Catharanthus roseus) cells, was isolated. Determination of the nucleotide sequence of the clone revealed that the cyc07 gene consists of seven exons separated by six introns. Genomic Southern analysis indicated that the cyc07 gene is present as a single copy per haploid genome in periwinkle. Expression of related genes was detected in a wide range of other plants. Transgenic Arabidopsis plants were generated that expressed the gene for β-glucuronidase (GUS) under the control of the promoter of the cyc07 gene. The tissue-specific pattern of expression directed by the promoter was investigated by analysis of GUS activity. Histochemical tests demonstrated that 589 bp of the 5′-upstream sequence of the cyc07 gene could direct specifical expression of the GUS reporter gene in meristematic tissues in transgenic plants. The spatial pattern of expression directed by the promoter was closely correlated with meristematic activity and cell proliferation, suggesting an association between the function of the cyc07 gene and cell proliferation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014441
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  • 42
    Electronic Resource
    Electronic Resource
    Over-production of the D1 protein of photosystem II reaction centre in the cyanobacterium Synechococcus sp. PCC 7942 (1994)
    Springer
    Plant molecular biology 26 (1994), S. 709-721 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; photosynthesis ; protein turnover ; psbA ; tac promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The unicellular cyanobacterium Synechococcus sp. PCC 7942 has three psbA genes encoding two different forms of the photosystem II reaction centre protein D1 (D1:1 and D1:2). The level of expression of these psbA genes and the synthesis of D1:1 and D1:2 are strongly regulated under varying light conditions. In order to better understand the regulatory mechanisms underlying these processes, we have constructed a strain of Synechococcus sp. PCC 7942 capable of over-producing psbA mRNA and D1 protein. In this study, we describe the over-expression of D1:1 using a tac-hybrid promoter in front of the psbAI gene in combination with lacI Q repressor system. Over-production of D1:1 was induced by growing cells for 12 h at 50 μmol photons m-2 s-1 in the presence of 40 or 80 μg/ml IPTG. The amount of psbAI mRNA and that of D1:1 protein in cells grown with IPTG was three times and two times higher, respectively. A higher concentration of IPTG (i.e., 150 μg/ml) did not further increase the production of the psbAI message or D1:1. The over-production of D1:1 caused a decrease in the level of D1:2 synthesised, resulting in most PSII reaction centres containing D1:1. However, the over-production of D1:1 had no effect on the pigment composition (chlorophyll a or phycocyanin/number of cells) or the light-saturated rate of photosynthesis. This and the fact that the total amounts of D1 and D2 proteins were not affected by IPTG suggest that the number of PSII centres within the membranes remained unchanged. From these results, we conclude that expression of psbAI can be regulated by using the tac promoter and lacI Q system. However, the accumulation of D1:1 protein into the membrane is regulated by the number of PSII centres.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00013756
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  • 43
    Electronic Resource
    Electronic Resource
    Environmental stress-mediated differential 3′ end formation of chloroplast RNA-binding protein transcripts (1994)
    Springer
    Plant molecular biology 26 (1994), S. 833-849 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; RNA stability regulation ; chloroplast RNA-binding protein (cRBP) ; environmental stress ; Mesembryanthemum crystallinum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the characterization of transcripts from the halophyte, Mesembryanthemum crystallinum, encoding a protein with high homology to chloroplast RNA-binding proteins (cRBP). In this plant chloroplast-related functions are largely protected against salt stress. cRBP transcripts are derived from a single gene, Mc32crbp, although three size classes of polyadenylated mRNAs are detected. Transcription rate and steady state amounts of mRNA are developmentally regulated and light controlled with strong transcriptional activity as functional chloroplasts are established, and with lower maintenance activity thereafter. Upon salt stress, the rate of transcription decreases, although transcript levels increase. Accompanying stress, a change in the distribution of transcript size classes is observed as the longest transcript with an untranslated 3′ end of 381 nucleotides increases relative to transcripts with shorter 3′ ends. The long transcript is characterized by the presence of five sequence elements in the 3′-untranslated region that are present in cRBP mRNAs from a variety of plants, although not all elements are found in each mRNA. The results may indicate a mechanism by which mRNA levels of constitutively light-regulated genes may be modulated without enhanced transcription in response to environmental cues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028852
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  • 44
    Electronic Resource
    Electronic Resource
    Expression of engineered antibodies in plant cells (1994)
    Springer
    Plant molecular biology 26 (1994), S. 1023-1030 
    Details
    ISSN: 1573-5028
    Keywords: immunoglobulin genes ; gene expression ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040685
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  • 45
    Electronic Resource
    Electronic Resource
    A method for examining expression of homologous genes in plant polyploids (1994)
    Springer
    Plant molecular biology 26 (1994), S. 1065-1071 
    Details
    ISSN: 1573-5028
    Keywords: Brassica ; polyploid ; gene expression ; RT-PCR ; RFLP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract One of the essential issues regarding evolution of polyploid species is how duplicate genes are expressed. Most studies on gene expression in polyploids have been based on isozyme analyses; RNA analysis has not been widely used partially due to difficulties in distinguishing homologous transcripts which usually have the same length and similar or almost identical sequences. In this study, a method combining RT-PCR with RFLP was used to analyze transcripts of homologous genes in natural and synthetic Brassica amphidiploids. Sequences coding for several known genes were selected and used to synthesize gene-specific primers. Total RNAs were used as templates for RT-PCR to amplify homologous transcripts in three diploid parental species, three cultivated amphidiploid species and six synthetic amphidiploids. For each gene, initial PCR products amplified in all species had identical length; however, homologous transcripts in the diploid and amphidiploid species could be distinguished after digesting the PCR products with restriction enzymes. Preliminary results based on three genes indicated that both transcripts from the diploid parents were expressed in the synthetic and natural amphidiploids. This study represents the first application of RT-PCR and RFLP analysis to investigate expression of homologous genes in higher plants. The technique is a sensitive, simple and efficient method for distinguishing homologous transcripts in a mixed RNA population and can be applied to many types of studies on expression of homologous genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040689
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  • 46
    Electronic Resource
    Electronic Resource
    Molecular cloning of the Arabidopsis thaliana sedoheptulose-1,7-biphosphatase gene and expression studies in wheat and Arabidopsis thaliana (1994)
    Springer
    Plant molecular biology 26 (1994), S. 1191-1200 
    Details
    ISSN: 1573-5028
    Keywords: Calvin cycle genes ; gene expression ; SBPase ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report here the isolation and nucleotide sequence of genomic clones encoding the chloroplast enzyme sedoheptulose-1,7-bisphosphatase (SBPase) from Arabidopsis thaliana. The coding region of this gene contains eight exons (72–76 bp) and seven introns (75–91 bp) and encodes a polypeptide of 393 amino acids. Unusually, the 5′ non-coding region contains two additional AUG codons upstream of the translation initiation codon. A comparison of the deduced Arabidopsis and wheat SBPase polypeptide sequences reveals 78.6%, identity. Expression studies showed that the level of SBPase mRNA in Arabidopsis and wheat is regulated in a light-dependent manner and is also influenced by the developmental stage of the leaf. Although the Arabidopsis SBPase gene is present in a single copy, two hybridizing transcripts were detected in some tissues, suggesting the presence of alternate transcription start sites in the upstream region.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040699
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  • 47
    Electronic Resource
    Electronic Resource
    Cotton (Gossypium hirsutum L.) pollen-specific polygalacturonase mRNA: tissue and temporal specificity of its promoter in transgenic tobacco (1994)
    Springer
    Plant molecular biology 26 (1994), S. 1989-1993 
    Details
    ISSN: 1573-5028
    Keywords: polygalacturonase ; pollen-specific promoter ; cotton ; transgenics ; transformation ; β-glucuronidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A gene (G9) expressed during late microsporogenesis in cotton (Gossypium hirsutum L.) was isolated. Sequence analysis of the cDNA (1.3 kb) as well as the gene (2.6 kb) revealed an open reading frame of 1233 bases encoding a protein of 43.9 kDa. The coding region of the gene is interrupted by three introns. Northern analysis of the RNA from developing anthers showed that the transcripts appear 12 days before anthesis and that the maximal concentration of RNA occurs in pollen on the day of anthesis. This pattern of gene expression suggests functions in post-anthesis events. Sequence comparisons with other known plant genes indicated that G9 is homologous to polygalacturonases. The G9 promoter conferred tissue and temporal specificity of β-glucuronidase (GUS) expression in transgenic tobacco plants. Thus, the G9 promoter can be used to drive gene expression in homologous as well as heterologous plants in a tissue-specific manner.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00019509
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  • 48
    Electronic Resource
    Electronic Resource
    Expression of the tobacco Tnt1 retrotransposon promoter in heterologous species (1994)
    Springer
    Plant molecular biology 26 (1994), S. 393-402 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; Brassica napus ; gene expression ; Nicotiana tabacum ; retrotransposon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of the tobacco (Nicotiana tabacum) retrotransposon Tntl has previously been shown to be strongly regulated and driven from the 5′ long terminal repeat (LTR). We report here that the Tntl LTR can promote activity of the β-glucuronidase (GUS) reporter gene in two heterologous species of the Brassicaceae family, namely rapessed (Brassica napus) and Arabidopsis thaliana. The translational LTR-GUS fusion was active in transient expression studies performed with tobacco and rapeseed protoplasts, indicating that the LTR sequences are recognized in heterologous species. Our results also showed that Tntl LTR-promoted GUS expression in transgenic Arabidopsis is strongly regulated, and that, in contrast to tobacco, hormonal activation plays a significant role in the expression of the Tntl LTR in Arabidopsis. LTR sequences were shown to be more effective than the CaMV 35S enhancer region in transient expression studies performed with tobacco or rapessed protoplasts; and substitution of the LTR sequences upstream from the major transcriptional start with the CaMV 35S enhancer region gave high levels of expression in transgenic tobacco and Arabidopsis leaves, suggesting that a Tntl element with similar substitutions in its 5′ LTR might be suited for gene-tagging experiments in heterologous species.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00039548
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  • 49
    Electronic Resource
    Electronic Resource
    Structure and promoter analysis of an ABA- and stress-regulated barley gene, HVA1 (1994)
    Springer
    Plant molecular biology 26 (1994), S. 617-630 
    Details
    ISSN: 1573-5028
    Keywords: ABA ; barley ; gene expression ; Hordeum vulgare ; phylogeny ; stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A single-copy barley gene, HVA1, encoding a class 3 late embryogenesis-abundant protein, can be induced by either treatment with abscisic acid (ABA) or by stress conditions such as drought, cold, heat and salinity. We have isolated an HVA1 genomic clone containing about 400 bp of 5′-upstream sequence, a single 109 bp intron, and the full coding sequence. Linker scan mutagenesis and transient expression studies were used to test the function of four HVA1 promoter elements conserved in ABA-responsive genes. Mutations in two of these elements, the C box and the putative ABRE 1 (ABA-responsive element) containing an ACGT core, resulted in no significant change in transcription level or ABA induction. In contrast, mutations of the other two elements, putative ABRE 2 & 3 cause the level of transcription to drop to 10–20% of that obtained with the wild-type promoter indicating that the high level of expression of HVA1 is dependent on both pABRE 2 & 3. Interestingly, despite their low level of expression, the mutated promoters still gave more than 20-fold induction in response to ABA treatment. We suggest that the ABA induction of barley HVA1 gene is governed by a complex consisting of pABRE 2 & 3 working together to regulate the absolute level of expression, and either of these elements or a possible third element may regulate ABA inducibility. Phylogenetic analysis by parsimony indicates that the barley HVA1 and wheat pMA2005 sequences share a recent common ancester. These two genes are closely related to the carrot Dc3 and cotton D-7 genes with which they share a similar structural gene organization.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00013748
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  • 50
    Electronic Resource
    Electronic Resource
    Detection of immunologically related Kunitz and Bowman-Birk proteinase inhibitors expressed during potato tuber development (1994)
    Springer
    Plant molecular biology 26 (1994), S. 961-969 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; Kunitz-type proteinase inhibitor ; potato (Solanum tuberosum, L.) ; soybean C-II inhibitor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Antiserum against a potato Kunitz-type proteinase inhibitor (PKPI) expressed in Escherichia coli was produced. In immunoblotting assays of proteins from potato tubers cultured in vitro, three proteins reacted to the antiserum, two of 20 kDa and one of 10 kDa. Their N-termini were sequenced. While the 20 kDa proteins showed 59 and 90% identity to PKPI, the 10 kDa one had 65% identity to soybean C-II proteinase inhibitor. Characterization of the temporal expression of these proteins showed that both could be detected from 10 days after induction of tuberization (DAI) in vitro, but the times when maximum amounts of PKPI and 10 kDa protein could be detected were different, corresponding to 22 and 32 DAI, respectively. The amounts of these proteins decreased in the following stages, and no positive reaction of the antiserum with mature tuber proteins could be found. The 20 kDa proteins were also detected in early stages of development of potato tubers grown in the field, indicating that these proteins are expressed during normal tuber development, and differ from the PKPIs reported previously.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028862
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  • 51
    Electronic Resource
    Electronic Resource
    Production of fertile transgenic maize by electroporation of suspension culture cells (1994)
    Springer
    Plant molecular biology 24 (1994), S. 51-61 
    Details
    ISSN: 1573-5028
    Keywords: Zea mays L. ; transformation ; electroporation ; bar ; phosphinothricin acetyltransferase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Fertile, transgenic maize plants were generated by electroporation of suspension culture cells that were treated with a pectin-degrading enzyme. Electroporation of cells from two different suspension cultures, one derived from A188 X B73 and one derived from a B73-related inbred, with a plasmid containing the bar gene, resulted in high-frequency recovery of stably transformed callus lines. Plants were regenerated from thirteen transformed callus lines and transmission of bar to progeny was demonstrated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040573
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  • 52
    Electronic Resource
    Electronic Resource
    Cloning and molecular analysis of structural genes involved in flavonoid and stilbene biosynthesis in grape (Vitis vinifera L.) (1994)
    Springer
    Plant molecular biology 24 (1994), S. 743-755 
    Details
    ISSN: 1573-5028
    Keywords: anthocyanins ; cDNA cloning ; flavonoids ; gene expression ; genomic organization ; stilbenes ; Vitis vinifera L
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genes involved in flavonoid and stilbene biosynthesis were isolated from grape (Vitis vinifera L.). Clones coding for phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydoxylase (F3H), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin dioxygenase (LDOX) and UDP glucose:flavonoid 3-O-glucosyl transferase (UFGT), were isolated by screening a cDNA library, obtained from mRNA from seedlings grown in light for 48 h using snapdragon (Antirrhinum majus) and maize heterologous probes. A cDNA clone coding for stilbene synthase (StSy) was isolated by probing the library with a specific oligonucleotide. These clones were sequenced and when the putative products were compared to the published amino acid sequence for corresponding enzymes, the percentages of similarity ranged from 65% (UFGT) to 90% (CHS and PAL). The analysis of the genomic organization and expression of these genes in response to light shows that PAL and StSy genes belong to large multigene families, while the others are present in one to four copies per haploid genome. The steady-state level of mRNAs encoded by the flavonoid biosynthetic genes as determined in young seedlings is coordinately induced by light, except for PAL and StSy, which appear to be constitutively expressed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00029856
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  • 53
    Electronic Resource
    Electronic Resource
    Triticum aestivum puroindolines, two basic cystine-rich seed proteins: cDNA sequence analysis and developmental gene expression (1994)
    Springer
    Plant molecular biology 25 (1994), S. 43-57 
    Details
    ISSN: 1573-5028
    Keywords: cDNA sequence ; cystine-rich proteins ; gene expression ; puroindolines ; tryptophan-rich domain ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract From a mid-maturation seed cDNA library we have isolated cDNA clones encoding two Triticum aestivum puroindolines. Puroindoline-a and puroindoline-b, which are 55% similar, are basic, cystine-rich and tryptophan-rich proteins. Puroindolines are synthezised as preproproteins which include N- and C-terminal propeptides which could be involved in their vacuolar localization. The mature proteins have a molecular mass of 13 kDa and a calculated isoelectric point greater than 10. A notable feature of the primary structure of puroindolines is the presence of a tryptophan-rich domain which also contains basic residues. A similar tryptophan-rich domain was found within an oat seed protein and a mammalian antimicrobial peptide. The ten cysteine residues of puroindolines are organized in a cysteine skeleton which shows similarity to the cysteine skeleton of other wheat seed cystine-rich proteins. Northern blot analysis showed that puroindoline genes are specifically expressed in T. aestivum developing seeds. No puroindoline transcripts as well as no related genes were detected in Triticum durum. The identity of puroindolines to wheat starch-granule associated proteins is discussed as well as the potential role of puroindolines in the plant defence mechanism.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00024197
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  • 54
    Electronic Resource
    Electronic Resource
    Structure of genes that encode isozymes of aspartate aminotransferase in Panicum miliaceum L., a C4 plant (1994)
    Springer
    Plant molecular biology 26 (1994), S. 723-734 
    Details
    ISSN: 1573-5028
    Keywords: aspartate aminotransferase ; C4 photosynthesis ; gene expression ; gene structure ; isozyme
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cytosolic and mitochondrial isozymes of aspartate aminotransferase (AspAT) function in the C4 photosynthetic cycle in NAD-malic enzyme-type C4 plants and are expressed at high levels in mesophyll cells and bundle sheath cells, respectively. We constructed a genomic library from Panicum miliaceum, a NAD-malic enzyme-type C4 plant, and cloned the genes for these isozymes. The sequence of the cloned gene for cytosolic AspAT spans 7800 bp and consists of 12 exons. The sequence of the cloned gene for mitochondrial AspAT spans 9000 bp and consists of 10 exons. The results of primer-extension analysis suggest that transcription may be initiated from multiple adjacent sites. Both genes have significant GC-rich regions around the site of initiation of transcription, and these regions showed no CpG suppression. The 5′-flanking regions of both genes include several short sequences similar to the regulatory elements found in other genes for components of the photosynthetic machinery. In particular, the cytosolic AspAT gene contains sequences similar to nuclear protein-binding sites in other mesophyll-expressed C4 photosynthetic genes and the mitochondrial AspAT gene contains elements for light-sensitive and constitutive expression of a bundle sheath-expressed gene. The results of Southern analysis indicated that there are at least two genes that encode each isozyme in the genome of P. miliaceum. A comparison of nitron-insertion positions between AspAT genes of plants and animals revealed that several introns are located at identical positions. On the basis of a phylogenetic tree among AspATs and tyrosine aminotransferase, we have shown that the introns of aminotransferase genes antedate the divergence of eubacteria, archaebacteria, and eukaryotes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00013757
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  • 55
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    Electronic Resource
    Vascular expression of the grp1.8 promoter is controlled by three specific regulatory elements and one unspecific activating sequence (1994)
    Springer
    Plant molecular biology 26 (1994), S. 747-756 
    Details
    ISSN: 1573-5028
    Keywords: activating sequence ; gene expression ; glycine-rich protein ; tobacco ; vascular expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The bean grp1.8 full-length promoter is specifically active in vascular tissue during normal development of tobacco. Deletion of a negative regulatory element resulted in ectopic activity of the promoter in cortical cells of hypocotyls, roots and stems. A 169 bp fragment (−205 to −36) of the grp1.8 promoter conferred vascular-specific expression to CaMV 35S minimal promoters whereas a 141 bp fragment (−205 to −64) strongly activated these minimal promoters both in vascular and cortical cells. These experiments defined a new regulatory element (VSE) that is essential for vascular-specific expression and is located between −64 and −36. The 141 bp grp1.8 promoter sequence had enhancer-like properties as it was active in both orientations. A 24 bp sequence (bp −119 to −96, corresponding to the SE1 regulatory element) enhanced expression from several minimal promoters strongly but unspecifically, whereas a 26 bp sequence (−98 to −73, corresponding to the RSE regulatory element) induced vascular-specific expression. Thus, the grp1.8 promoter is regulated by a combinatorial mechanism that can integrate the action of different, non-additively acting regulatory elements into vascular-specific expression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00013759
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    A dehydrin cognate protein from pea (Pisum sativum L.) with an atypical pattern of expression (1994)
    Springer
    Plant molecular biology 26 (1994), S. 805-816 
    Details
    ISSN: 1573-5028
    Keywords: Dehydrin ; gene expression ; pea (Pisum sativum L.) ; cognate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Dehydrins are a family of proteins characterised by conserved amino acid motifs, and induced in plants by dehydration or treatment with ABA. An antiserum was raised against a synthetic oligopeptide based on the most highly conserved dehydrin amino acid motif, the lysine-rich block (core sequence KIKEK-LPG). This antiserum detected a novel M r 40 000 polypeptide and enabled isolation of a corresponding cDNA clone, pPsB61 (B61). The deduced amino acid sequence contained two lysine-rich blocks, however the remainder of the sequence differed markedly from other pea dehydrins. Surprisingly, the sequence contained a stretch of serine residues, a characteristic common to dehydrins from many plant species but which is missing in pea dehydrin. The expression patterns of B61 mRNA and polypeptide were distinctively different from those of the pea dehydrins during seed development, germination and in young seedlings exposed to dehydration stress or treated with ABA. In particular, dehydration stress led to slightly reduced levels of B61 RNA, and ABA application to young seedlings had no marked effect on its abundance. The M r 40 000 polypeptide is thus related to pea dehydrin by the presence of the most highly conserved amino acid sequence motifs, but lacks the characteristic expression pattern of dehydrin. By analogy with heat shock cognate proteins we refer to this protein as a dehydrin cognate.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028850
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    Gibberellins: perception, transduction and responses (1994)
    Springer
    Plant molecular biology 26 (1994), S. 1529-1555 
    Details
    ISSN: 1573-5028
    Keywords: gibberellin ; growth ; development ; perception ; receptor ; gene expression ; signal transduction ; response mutant ; calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00016489
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    Isolation and characterization of two β-tubulin cDNA clones from rice (1994)
    Springer
    Plant molecular biology 26 (1994), S. 1975-1979 
    Details
    ISSN: 1573-5028
    Keywords: β-tubulin ; cDNA ; rice ; monocot ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two cDNA clones encoding two different β-tubulins, RTUB-1 and RTUB-2, were isolated from a rice cDNA library and their nucleotide sequences were analyzed. The deduced amino acid sequences showed amino acid sequence identity between 92% and 97% with other plant β-tubulins. Southern blot analysis using gene-specific and coding-region probes suggested that β-tubulins in rice are encoded by multigene families. The two cDNA clones represent two subfamilies of rice tubulins. RTUB-1 and RTUB-2, consisting of 3 to 4 genes and a single gene, respectively. The transcript levels of RTUB-1 and RTUB-2 genes were higher in actively elongating tissues such as etiolated shoot tissues and light-grown root tissues of four-day old seedlings.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00019507
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  • 59
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    Hairy roots of Brassica napus: II. Glutamine synthetase overexpression alters ammonia assimilation and the response to phosphinothricin (1994)
    Springer
    Plant cell reports 14 (1994), S. 41-46 
    Details
    ISSN: 1432-203X
    Keywords: Agrobacterium rhizogenes ; Brassica napus ; glutamine synthetase ; phosphinothricin ; rape ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Hairy roots of Brassica napus (rape cv. Giant) have been produced that contain the cytosolic glutamine synthetase (GS) gene from Glycine max (soybean). Leaf explants were cocultivated with Agrobacterium rhizogenes strain A4T harbouring the binary vector pLN16. This vector was constructed by inserting a soybean cytosolic GS cDNA into the multiple cloning site of pGA643, placing it under the control of the CaMV promoter. In addition, the T-DNA region of pLN16 contained a NPTII gene for selection of transformed cells. Transgenic hairy roots grew prolifically on hormone-free media containing a selective level of kanamycin. Southern and northern analyses confirmed the presence of soybean GS DNA and transcripts, respectively. These transformed hairy roots also have a greater abundance of the GS polypeptide, approximately 3–6 fold greater GS activity and lower levels of endogenous ammonia. Hairy roots provide a useful system for studying responses to phosphinothricin (PPT). Hairy roots grown in media containing PPT had lower GS activity, greater ammonia accumulation and slower growth than controls. The presence of the soybean GS gene in the hairy roots reduced these PPT-induced effects and resulted in higher GS activity, lower ammonia levels and faster growth than in PPT-treated controls. Greater tolerance of PPT was also seen in shoots regenerated from the hairy roots displaying elevated levels of GS activity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233296
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    Changes in polypeptide patterns in tobacco roots colonized by two Glomus species (1994)
    Springer
    Mycorrhiza 4 (1994), S. 215-221 
    Details
    ISSN: 1432-1890
    Keywords: Nicotiana ; Glomus species ; arbuscular mycorrhiza ; gene expression ; specific polypeptides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Changes in gene expression were studied during the establishment of arbuscular mycorrhizal symbiosis in tobacco roots from an amphidiploid hybrid Nicotiana glutinosa x N. debneyi. Polypeptide patterns from control roots and from roots infected by Glomus mosseae or G. intraradices were resolved by two-dimensional polyacrylamide gel electrophoresis and followed in a time-course analysis. Arbuscular mycorrhizal infection led to significant modifications in polypeptide patterns with: (a) decreased amounts of some polypeptides, (b) increased accumulation of others, and (c) appearance of newly-induced polypeptides. Comparisons made during infection development by the two Glomus species demonstrated that protein modifications changed in relation to the mycorrhizal state of the tobacco roots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00206783
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    T-DNA-insert-independent mutations induced in transformed plant cells duringAgrobacterium co-cultivation (1994)
    Springer
    Transgenic research 3 (1994), S. 317-325 
    Details
    ISSN: 1573-9368
    Keywords: Agrobacterium ; mutagenesis ; Nicotiana plumbaginifolia ; nitrate reductase ; ploidy ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transformation frequencies were determined for 1n, 2n, and 4n Nicotiana plumbaginifolia protoplast cultures inAgrobacterium-mediated gene transfer experiments. An unexpected large drop (50%) in plating efficiencies was observed in the non-selected (control) 1n populations after transformation treatment with virulent strains. This effect was not observed in the 2n or 4n cultures or in the 1n cultures when treated with avirulent bacteria. The mortality was disproportionally high and could not be explained by the low (0.1–0.5%) transformation efficiency in the 1n population, indicating mutagenesis of the cell populations independently from the T-DNA insertions. Mutagenesis was also indicated in gene tagging experiments where nitrate reductase-deficient (NR−) mutants were selected from haploidNicotiana plumbaginifolia protoplasts, as well as from leaf disc cultures or protoplasts of diploid plants that were heterozygotic for a mutation either in the NR apoenzyme gene (nia/wt) or one of the molybdenum-containing cofactor genes (cnxA/wt), afterAgrobacterium co-cultivation. The chlorate-resistant isolates were tested for the T-DNA-specific kanamycin resistance trait only after NR-deficiency had been established. Thirty-nine independent NR-deficient mutants were analysed further by Southern blot hybridization. There was no indication of integrated T-DNA sequences in the mutated NR genes, despite the fact that NR-deficient cells were found more frequently in cell populations which became transformed during the treatment than in the populations which did not. These observations suggest that transformation-competent cells undergo mutagenesis during theAgrobacterium gene transfer process not only as a result of stable integration events, but also through accompanying events that do not result in major changes in the mutated loci. The nature of these changes at the molecular level remains to be elucidated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01973592
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    Tissue specific expression of an α-skeletal actin-lacZ fusion gene during development in transgenic mice (1994)
    Springer
    Transgenic research 3 (1994), S. 59-66 
    Details
    ISSN: 1573-9368
    Keywords: α-actin ; transgenic mice ; gene expression ; muscle ; embryos ; lacZ
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic mice carrying a chimaeric transgene containing 730 bp of the 5′-flanking sequences and the entire first intron of the rat α-skeletal actin gene fused to thelacZ reporter gene have been produced by microinjection. ThelacZ reporter gene was used to verify the suitability of using the rat α-actin promoter elements to target expression of genes of agricultural and therapeutic value exclusively to skeletal and heart muscle cells and fibres of transgenic mice. Expression of the transgene indicates a tightly regulated developmental and muscle specific control of the rat α-skeletal actin gene, making it a useful promoter for gene targeting to muscle tissues. The cells destined to form muscle tissues in these transgenic mice are readily visualized in intact embryos by staining for β-galactosidase activity, making them a suitable animal model for studying the origin and development of skeletal and cardiac muscle tissues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01976028
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    An assessment of gene transfer by pollen from field-grown transgenic potatoes to non-transgenic potatoes and related species (1994)
    Springer
    Transgenic research 3 (1994), S. 216-225 
    Details
    ISSN: 1573-9368
    Keywords: Solanum tuberosum ; genetic modification ; transformation ; gene transfer ; genetic isolation ; risk assessment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Information on the extent of transgene dispersal by pollen to adjacent potato plots and to related weed species is an important requisite for risk assessment; a procedure followed before novel transgenic plants are evaluated under field conditions. The purpose of the investigation was to determine the frequency of cross-pollination between potato (Solanum tuberosum) plants at different distances, using a kanamycin resistnace transgene (nptII) as a selectable marker. All potato plants were from the variety Désirée. Non-transgenic potato plants, used as potential recipients of transgene-containing pollen, were planted in 12 sub-plots, at distances of 0–20 m from the nearest transgenic potato plants. Seeds harvested from the non-transgenic plants were screened for resistance to kanamycin, and molecular methods were used to confirm that resistant progeny contained thenptII gene. Where transgenic and non-transgenic potato plants were in alternate rows (leaves touching), 24% of seedlings from the non-transgenic parent plants were kanamycin-resistant. Comparable seedlings from plants at up to 3 m distance had a resistance frequency of 2%, at 10 m the frequency was 0.017% and at 20 m no resistant progeny were observed. Plants of the weed speciesS. dulcamara andS. nigrum were also planted close to the transgenic potatoes to test for evidence of hybridization, and no kanamycin-resistant seedlings were observed among progeny fromS. dulcamara andS. nigrum. This investigation provided evidence that the extent of gene dispersal from transgenic potatoes to non-transgenic potatoes falls markedly with increasing distance, and is negligible at 10 m. There was, also, no evidence of transgene movement from potato toS. dulcamara andS. nigrum under field conditions. These data will be valuable in defining genetic isolation procedures for the early field evaluation and the use of novel transgenic potato genotypes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02336774
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    Complementation of an alcohol dehydrogenase-deficientNicotiana plumbaginifolia mutant by transformation with theArabidopsis thaliana Adh gene (1994)
    Springer
    Transgenic research 3 (1994), S. 376-387 
    Details
    ISSN: 1573-9368
    Keywords: alcohol dehydrogenase ; Arabidopsis thaliana ; functional complementation ; gene expression ; Nicotiana plumbaginifolia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An alcohol dehydrogenase-deficient (ADH) mutant ofNicotiana plumbaginifolia, selected on the basis of ethanol resistance, was restored for ADH activity by transformation with anAdh gene fromArabidopsis thaliana expressed under the control of its own promoter or the CaMV 35S promoter. The expression in various organs (seed, root, leaf and pollen) was analysed at the protein and RNA levels as well as byin situ detection of ADH activity. The analysis of spatial and temporal regulation of theA. thaliana Adh gene expression suggests that ADH expression is controlled at the transcriptional level.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01976769
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    Recombinant viruses as transformation vectors of marine macroalgae (1994)
    Springer
    Journal of applied phycology 6 (1994), S. 247-253 
    Details
    ISSN: 1573-5176
    Keywords: algae ; genes ; recombinant ; transformation ; vectors ; viruses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The large dsDNA viruses that are known to infect eukaryotic algae show promise as genetic vectors for algal biotechnology. The large size (150–330 kbp) of these viral genomes may permit insertion of large sequences of foreign DNA. The viruses infecting filamentous marine brown algae appear to be integrated into the genomes of their hosts, and may provide integration mechanisms that can be used for directing insertion of foreign genes into algal chromosomes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02186078
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    Carbohydrate binding activities ofBradyrhizobium japonicum: IV. Effect of lactose and flavones on the expression of the lectin, BJ38 (1994)
    Springer
    Glycoconjugate journal 11 (1994), S. 363-370 
    Details
    ISSN: 1573-4986
    Keywords: lectin ; gene expression ; cell-cell adhesion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract BJ38 is a galactose/lactose-specific lectin (M r ∼ 38000) found at one pole ofBradyrhizobium japonicum. It has been implicated in mediating the adhesion of the bacteria to soybean roots, leading to the establishment of a nitrogen-fixing symbiosis. When the ligand lactose is added to cultures of the bacteria for at least 1 h prior to harvesting the cells for BJ38 isolation, the yield of the protein was found to be elevated in a dose-dependent fashion. Half maximal stimulation was observed at ∼ 50 µm; the effect was saturated at ∼ 1mm, where a 10-fold higher yield of BJ38 was obtained. Saccharides with a lower affinity for BJ38 than lactose yielded a correspondingly smaller induction effect when compared at a concentration of 1mm. The higher level of BJ38 induced by lactose is also manifested by an elevated amount of BJ38 detectable at the cell surface and by a higher number ofB. japonicum cells adsorbed onto soybean cells. Surprisingly, the induction of BJ38 expression seen with lactose was also observed with certain, but not all, flavonoids that induce thenod genes of the bacteria; genistein mimicked the induction observed with lactose, whereas luteolin failed to stimulate BJ38 production.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00731210
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    Recombinant protein expression in aDrosophila cell line: comparison with the baculovirus system (1994)
    Springer
    Cytotechnology 15 (1994), S. 139-144 
    Details
    ISSN: 1573-0778
    Keywords: Baculovirus ; cell culture ; Drosophila ; gene expression ; insect cell ; metallothionein promoter ; recombinant protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In this report, we compare two different expression systems: baculovirus/Sf9 and stable recombinantDrosophila Schneider 2 (S2) cell lines. The construction of a recombinant S2 cell line is simple and quick, and in batch fermentations the cells have a doubling time of 20 hours until reaching a plateau density of 20 million cells/ml. Protein expression is driven by theDrosophila Metallothionein promoter which is tightly regulated. When expressed in S2 cells, the extracellular domain of human VCAM, an adhesion molecule, is indistinguishable from the same protein produced by baculovirus-infected Sf9 cells. Additionally, we present data on the expression of a seven trans-membrane protein, the dopamine D4 receptor, which has been successfully expressed in both systems. The receptor integrates correctly in the S2 membrane, binds [3H]spiperone with high affinity and exhibits pharmacological characteristics identical to that of the receptor expressed in Sf9 and mammalian cells. The general implications for large scale production of recombinant proteins are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00762388
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    Maintenance and expression of heterologous genes in chloroplast ofChlamydomonas reinhardtii (1994)
    Springer
    Journal of applied phycology 6 (1994), S. 239-245 
    Details
    ISSN: 1573-5176
    Keywords: Chlamydomonas reinhardtii ; transformation ; chloroplast ; aminoglycoside adenine transferase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The chloroplast genome ofChlamydomonas reinhardtii has been transformed with a chimeric gene consisting of the chloroplastatpA promoter and the bacterial gene for aminoglycoside adenine transferase (aadA). TheatpA-aadA cassette has been placed within the chloroplast DNAEcoRI restriction enzyme fragment 14, or within the chloroplastBamH1 fragment 10. The chimeric constructs were introduced into the chloroplast by particle bombardment. Integration of the cassette into chloroplast DNA then occurred via homologous recombination of sequences flanking the cassette with their corresponding chloroplast sequences. We demonstrate that the chloroplastatpA promoter inatpA-aadA routinely recombines with its endogenous counterpart, resulting in heteroplasmic chloroplast DNA populations that may persist for many generations. The heterologous gene does not require a 3′ inverted repeat sequence for its expression. TheatpA-aadA gene copy number, which is dictated here by its position in the chloroplast genome, is proportional to the steady state level ofatpA-aadA mRNA. However, neither genomic position, gene copy number, or mRNA level have a significant effect on cellular resistance to spectinomycin, nor activity of theaadA gene productin vitro. These results suggest that, in the case ofaadA, the limiting step for expression of this gene is at the translational or post-translational level. TheatpA-aadA cassette should prove a useful model for future studies on the maintenance and expression of heterologous genes inC. reinhardtii chloroplasts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02186077
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    A simple technique for producing 13C or 14C-labelled fronds of Lemna gibba and their use in soil incubation investigations (1994)
    Springer
    Plant and soil 160 (1994), S. 185-191 
    Details
    ISSN: 1573-5036
    Keywords: carbon-labelling ; carbon dioxide production ; decomposition ; 14C-glucose ; Lemna ; soil organic matter ; sugars ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The duckweed Lemna gibba required light and a suitable energy source such as sucrose, glucose or fructose, for maximum growth in culture. The requirement for light was relatively unimportant and the plants grew well in a photon flux density of only 52 μmol m-2s-1 PAR. The uptake and incorporation of uniformly labelled 14C-glucose into fronds was related only to the concentration of the sugar. When incubated with soil, labelled L. gibba behaved in a manner similar to that of labelled ryegrass roots which had been produced by a more elaborate technique using a 14CO2 labelled atmosphere. During incubation with soil for 224 days the L. gibba material (specific activity 6133 Bq mg-1 d. wt) lost 64% of its radioactivity as 14CO2 and ryegrass (specific activity 6634 Bq mg-1 d. wt) lost 49%. Alkaline extracted humic and fulvic acids from soil had specific activities for the L. gibba incubation of 3409 and 407 Bq mg-1 solid and for ryegrass roots of 4609 and 546 Bq mg-1 solid respectively. The production of 13C or 14C-labelled L. gibba can be undertaken using only simple equipment producing material the specific radioactivity of which can be controlled by adjusting the activity of the sugar energy source.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00010144
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    UV-B damage and protection at the molecular level in plants (1994)
    Springer
    Photosynthesis research 39 (1994), S. 475-489 
    Details
    ISSN: 1573-5079
    Keywords: DNA repair ; flavonoids ; gene expression ; oxidative stress ; photosynthesis ; promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Influx of solar UV-B radiation (280–320 nm) will probably increase in the future due to depletion of stratospheric ozone. In plants, there are several targets for the deleterious UV-B radiation, especially the chloroplast. This review summarizes the early effects and responses of low doses of UV-B at the molecular level. The DNA molecules of the plant cells are damaged by UV due to the formation of different photoproducts, such as pyrimidine dimers, which in turn can be combatted by specialized photoreactivating enzyme systems. In the chloroplast, the integrity of the thylakoid membrane seems to be much more sensitive than the activities of the photosynthetic components bound within. However, the decrease of mRNA transcripts for the photosynthetic complexes and other chloroplast proteins are among very early events of UV-B damage, as well as protein synthesis. Other genes, encoding defence-related enzymes, e.g., of the flavonoid biosynthetic pathway, are rapidly up-regulated after commencement of UV-B exposure. Some of the cis-acting nucleotide elements and trans-acting protein factors needed to regulate the UV-induced expression of the parsley chalcone synthase gene are known.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014600
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    A robust protocol for site-directed mutagenesis of the D1 protein inChlamydomonas reinhardtii: A PCR-splicedpsbA gene in a plasmid conferring spectinomycin resistance was introduced into apsbA deletion strain (1994)
    Springer
    Photosynthesis research 42 (1994), S. 121-131 
    Details
    ISSN: 1573-5079
    Keywords: chloroplast ; Photosystem II ; psbA ; site-directed mutagenesis ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this paper, we describe a protocol to obtain a site-directed mutants in thepsbA gene ofChlamydomonas reinhardtii, which overcomes several drawbacks of previous protocols, and makes it possible to generate a mutant within a month. Since the large size of the gene, and the presence of four large introns has made molecular genetics of thepsbA gene rather unwieldy, we have spliced all of the exons of thepsbA gene by PCR to facilitate genetic manipulation and sequencing of the gene. The resultant construct (plasmid pBA153, with several unique restriction sites introduced at exon boundaries) carried 1.2 and 1.8 kb intact sequences from the 5′- and 3′-flanking regions, respectively. The plasmid was used to transform a D1-deletion mutant and was found to complement the deletion and restore photosynthetic activity. In addition, a bacterialaadA gene conferring spectinomycin resistance (spe r) was inserted downstream of the intron-freepsbA gene, to give construct pBA155. This allowed selection of mutant strains deficient in photosynthesis by using spectinomycin resistance, and eliminated the possibility of selection for revertant strains which is a consequence of having to use photosynthetic activity as a selection pressure. Finally, pBA155 was used to construct pBA157, in which additional restriction sites were inserted to facilitate cassette mutagenesis for generation of mutations in spans thought to be involved in donor-side interactions. AllpsbA deletion strains transformed with intron-freepsbA-aadA constructs encoding the wild-type D1 sequence, and screened on spectinomycin plates for thespe r phenotype, were able to grow photosynthetically, and all showed identical kinetics for electron transfer from primary (QA) to secondary quinone (QB) in Photosystem II, as assayed by the decay of the high fluorescence yield on oxidation of the reduced primary acceptor (QA −).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02187123
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    Acclimation of photosynthetic proteins to rising atmospheric CO2 (1994)
    Springer
    Photosynthesis research 39 (1994), S. 413-425 
    Details
    ISSN: 1573-5079
    Keywords: elevated CO2 ; gene expression ; Rubisco ; rbcL ; rbcS
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this review we discuss how the photosynthetic apparatus, particularly Rubisco, acclimates to rising atmospheric CO2 concentrations (ca). Elevated ca alters the control exerted by different enzymes of the Calvin cycle on the overall rate of photosynthetic CO2 assimilation, so altering the requirement for different functional proteins. A decreased flux of carbon through the photorespiratory pathway will decrease requirements for these enzymes. From modeling of the response of CO2 uptake (A) to intracellular CO2 concentration (ci) it is shown that the requirement for Rubisco is decreased at elevated ca, whilst that for proteins limiting ribulose 1,5 bisphosphate regeneration may be increased. This balance may be altered by other interactions, in particular plasticity of sinks for photoassimilate and nitrogen supply; hypotheses on these interactions are presented. It is speculated that increased accumulation of carbohydrate in leaves developed at elevated ca may signal the ‘down regulation’ of Rubisco. The molecular basis of this ‘down regulation’ is discussed in terms of the repression of photosynthetic gene expression by the elevated carbohydrate concentrations. This molecular model is then used to predict patterns of acclimation of perennials to long term growth in elevated ca.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014595
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    Mitochondrial DNA diseases: Histological and cellular studies (1994)
    Springer
    Journal of bioenergetics and biomembranes 26 (1994), S. 301-310 
    Details
    ISSN: 1573-6881
    Keywords: Mitochondrial encephalomyopathy ; mitochondrial DNA ; gene expression ; protein translation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Large-scale deletions and tRNA point mutations in mitochondrial DNA (mtDNA) are associated with a variety of different mitochondrial encephalomyopathies. Skeletal muscle in these patients shows a typical pathology, characterized by the focal accumulation of large numbers of morphologically and biochemically abnormal mitochondria (ragged-red fibers). Both mtDNA deletions and tRNA point mutations impair mitochondrial translation and produce deficiencies in oxidative phosphorylation. However, mutant and wild-type mtDNAs co-exist (mtDNA heteroplasmy) and the translation defect is not expressed until the ratio of mutant: wild-type mtDNAs exceeds a specific threshold. Below the threshold the phenotype can be rescued by intramitochondrial genetic complementation. The mosaic expression of the skeletal muscle pathology is thus determined by both the cellular and organellar distribution of mtDNA mutants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00763101
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    PC12 variants deficient in norepinephrine transporter mRNA have wild type activities of several other related transporters (1994)
    Springer
    Neurochemical research 19 (1994), S. 743-751 
    Details
    ISSN: 1573-6903
    Keywords: Neurotransmitters ; reuptake ; PC12 ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Wild type PC12 pheochromocytoma cells express a Na+-dependent norepinephrine transporter that operates in the uptake of catecholamines. In addition to the previously described Na+-dependent system A for the uptake of α-amino-isobutyric acid and system Gly for glycine, we have identified two other Na+-dependent transporter systems for amino acid uptake in these cells: 1) system β for β-alanine and taurine; and 2) a system for creatine. Uptake of α-amino-isobutyric acid, glycine, β-alanine, and creatine is not affected in some PC12 variants that were previously shown to be deficient in catecholamine uptake and to have decreased levels of norepinephrine transporter mRNA. We have isolated two PC12 cDNA clones that are essentially identical in sequence to recently reported cDNAs for rat brain taurine and creatine transporters, respectively, and a third cDNA that appears to code for a novel transporter. mRNAs for these three transporters are present at wild type levels in those variants that express no or little norepinephrine transporter mRNA. These results support the notion that the expression of catecholamine reuptake transporters may be particularly susceptible to down-regulation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00967715
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  • 75
    Electronic Resource
    Electronic Resource
    Mechanisms of EGF receptor regulation in breast cancer cells (1994)
    Springer
    Breast cancer research and treatment 31 (1994), S. 227-236 
    Details
    ISSN: 1573-7217
    Keywords: breast cancer ; epidermal growth factor receptor ; gene expression ; hormone-indepence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Overexpression of the EGF receptor in breast cancer correlates with poor prognosis and failure on endocrine therapy for both ER−/EGFR+ and ER+/EGFR+ tumors, suggesting a role for EGFR in the progression to hormone independence. The identification of specific DNase I hypersensitive site patterns for the EGFR gene in ER+ vs. ER− cells implicates regions of the EGFR first intron in up-regulation of EGFR, while estrogen regulation studies indicate the involvement of a repressor(s) in the maintenance of low levels of EGFR. Based on these findings, a multi-step model is proposed for the progression of breast cancer from a hormone-dependent, ER+/EGFR- phenotype to an aggressive, hormone-independent, ER−/EGFR+ stage.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00666156
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  • 76
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    Electronic Resource
    Some features about transposition of the maize elementDissociation inNicotiana plumbaginifolia (1994)
    Springer
    Genetica 93 (1994), S. 41-48 
    Details
    ISSN: 1573-6857
    Keywords: Ac/Ds ; transformation ; transgenic plants ; transposon tagging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have recently shown that a plasmid-borneDissociation (Ds) element can excise from extrachromosomal plasmid DNA and integrate into a plant genome in the presence of theActivator (Ac) transposase.Ds andAc-carrying plasmids were used to co-transformNicotiana plumbaginifolia protoplasts. Transgenic plants were regenerated and analyzed. Here we describe further characterization of the system and discuss its efficiency in terms of DNA transformation and transposon tagging.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01435238
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  • 77
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    Effect of a new cholecystokinin antagonist (FK 480) on gene expression of cholecystokinin and secretin in rat intestine (1994)
    Springer
    Journal of gastroenterology 29 (1994), S. 385-387 
    Details
    ISSN: 1435-5922
    Keywords: CCK antagonist (FK 480) ; gene expression ; CCK ; secretin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effects of a new cholecystokinin (CCK) antagonist (FK 480; 0.1 mg/kg per day given by intragastric administration to rats for 3 days) on the expression of the CCK and secretin genes, plasma CCK immunoreactivity, and CCK content in the intestinal mucosa were examined. FK 480 increased the level of CCK mRNA in the intestine to 1.7 times the level in control rats, but did not affect the level of secretin mRNA. It did not increase plasma CCK immunoreactivity or CCK content in the intestinal mucosa. These results suggest that the ingested FK 480 directly increased CCK mRNA level in the intestine and produced a dissociation between the synthesis and release of CCK.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02358383
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  • 78
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    Transmission of injected DNA sequences to multiple eggs of Metaseiulus occidentalis and Amblyseius finlandicus (Acari: Phytoseiidae) following maternal microinjection (1994)
    Springer
    Experimental and applied acarology 18 (1994), S. 319-330 
    Details
    ISSN: 1572-9702
    Keywords: Maternal microinjection ; transformation ; genetic improvement
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The persistence of DNA injected into two species of adult female phytoseiids and its transmission to serial eggs deposited by them was assessed by the polymerase chain reaction (PCR). The effect of DNA concentration on persistence and transmission was examined in Metaseiulus occidentalis. M. occidentalis females were microinjected with plasmid DNA at three different concentrations (250, 500, 750 ng μL−1) and allowed to deposit one to five eggs before the females and their last eggs were analyzed. Plasmid DNA was found in 82% of the females assayed and in 70% of all the eggs analyzed (including the fifth eggs produced after microinjection). Transmission of DNA to multiple eggs was also examined in Amblyseius finlandicus. Females of this species are less traumatized by microinjection allowing analysis of transmission over a more extended number of eggs. Females were microinjected and allowed to deposit eggs until their death. DNA from every fifth egg was analyzed by the PCR. PCR products were amplified from 51% of the eggs and from all egg classes except the 30th egg. The persistence and presence of plasmid DNA in both eggs and females suggests that (1) maternal microinjection is a more efficient method for DNA delivery than traditional egg microinjection, (2) it may be possible to isolate transformants from fewer maternally-microinjected females than originally expected, and (3) maternal microinjection could be useful as a DNA delivery system in other phytoseiids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00116313
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  • 79
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    Electronic Resource
    Photosynthetic electron transport and anaerobic metabolism in purple non-sulfur phototrophic bacteria (1994)
    Springer
    Antonie van Leeuwenhoek 66 (1994), S. 151-164 
    Details
    ISSN: 1572-9699
    Keywords: purple non-sulfur bacteria ; Rhodobacter ; photosynthesis ; CO2 fixation ; anaerobic respiration ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Purple non-sulfur phototrophic bacteria, exemplifed byRhodobacter capsulatus andRhodobacter sphaeroides, exhibit a remarkable versatility in their anaerobic metabolism. In these bacteria the photosynthetic apparatus, enzymes involved in CO2 fixation and pathways of anaerobic respiration are all induced upon a reduction in oxygen tension. Recently, there have been significant advances in the understanding of molecular properties of the photosynthetic apparatus and the control of the expression of genes involved in photosynthesis and CO2 fixation. In addition, anaerobic respiratory pathways have been characterised and their interaction with photosynthetic electron transport has been described. This review will survey these advances and will discuss the ways in which photosynthetic electron transport and oxidation-reduction processes are integrated during photoautotrophic and photoheterotrophic growth.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00871637
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  • 80
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    Genetics and gibberellin production inGibberella fujikuroi (1994)
    Springer
    Antonie van Leeuwenhoek 65 (1994), S. 217-225 
    Details
    ISSN: 1572-9699
    Keywords: Gibberella fujikuroi ; gibberellins ; mutants ; regulation by nitrogen ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Gibberella fujikuroi (Fusarium moniliforme) is a complex group of plant pathogens. Some strains produce gibberellic acid and other gibberellins that promote growth and regulate various stages in plant development. The paper describes the research effort directed to development of genetic tools for this species. Furthermore the main features of the gibberellin biosynthetic pathway as established in Gibberella are described.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00871950
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  • 81
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    Electronic Resource
    Strategies for improving heterologous protein production from filamentous fungi (1994)
    Springer
    Antonie van Leeuwenhoek 65 (1994), S. 245-250 
    Details
    ISSN: 1572-9699
    Keywords: Aspergillus ; gene expression ; heterologous protein ; protein secretion ; Trichoderma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Despite the naturally high capacity for protein secretion by many species of filamentous fungi, secteted yields of many heterologous proteins have been comparatively low. The strategies for yield improvement have included the use of strong homologous promoters, increased gene copy number, gene fusions with a gene encoding a naturally well-secreted protein, protease-deficient host strains and screening for high yields following random mutagenesis. Such approaches have been effective with some target heterologous proteins but not others. Approaches used in heterologous protein production from filamentous fungi are discussed and a perspective on emerging strategies is presented.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00871952
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  • 82
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    Electronic Resource
    Photobiology of Bacteria (1994)
    Springer
    Antonie van Leeuwenhoek 65 (1994), S. 331-347 
    Details
    ISSN: 1572-9699
    Keywords: photoactive proteins ; photoreceptors ; chromophores ; energy transduction ; light signalling ; phototaxis ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The field of photobiology is concerned with the interactions between light and living matter. For Bacteria this interaction serves three recognisable physiological functions: provision of energy, protection against excess radiation and signalling (for motility and gene expression). The chemical structure of the primary light-absorbing components in biology (the chromophores of photoactive proteins) is surprisingly simple: tetrapyrroles, polyenes and derivatised aromats are the most abundant ones. The same is true for the photochemistry that is catalysed by these chromophores: this is limited to light-induced exciton- or electron-transfer and photoisomerization. The apoproteins surrounding the chromophores provide them with the required specificity to function in various aspects of photosynthesis, photorepair, photoprotection and photosignalling. Particularly in photosynthesis several of these processes have been resolved in great detail, for others at best only a physiological description can be given. In this contribution we discuss selected examples from various parts of the field of photobiology of Bacteria. Most examples have been taken from the purple bacteria and the cyanobacteria, with special emphasis on recently characterised signalling photoreceptors inEctothiorhodospira halophila and inFremyella diplosiphon.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00872217
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  • 83
    Electronic Resource
    Electronic Resource
    Powdery mildew resistance in Aegilops tauschii coss. and synthetic hexaploid wheats (1994)
    Springer
    Genetic resources and crop evolution 41 (1994), S. 151-158 
    Details
    ISSN: 1573-5109
    Keywords: Triticum aestivum ; Aegilops tauschii (syn. Ae. squarrosa) ; Erysiphe graminis f. sp. tritici resistance genes ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A collection of 400 Ae. tauschii (syn. Ae. squarrosa) Coss. accessions were screened for powdery mildew resistance based on the response patterns of 13 wheat cultivars/lines possessing major resistance genes to nine differential mildew isolates. 106 accessions showed complete resistance to all isolates, and 174 accessions revealed isolate-specific resistance, among which were 40 accessions exhibiting an identical response pattern as wheat cultivar ‘Ulka/*8Cc’ which is known to possess resistance gene Pm2. Expression of both complete and isolate-specific resistance from Ae. tauschii was observed in some synthetic hexaploid wheats derived from four mildew susceptible T. durum Desf. parents, each crossed with five to 38 resistant diploid Ae. tauschii accessions. Synthetic amphiploids involving different combinations of T. durum and Ae. tauschii generally showed a decrease in resistance compared with that expressed by the Ae. tauschii parental lines.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00051631
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  • 84
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    Molecular biology of C4 phosphoenolpyruvate carboxylase: Structure, regulation and genetic engineering (1994)
    Springer
    Photosynthesis research 39 (1994), S. 115-135 
    Details
    ISSN: 1573-5079
    Keywords: C4 photosynthesis ; gene expression ; oligomerization ; phosphorylation-dephosphorylation cascade ; PEPC-protein kinase ; site-directed mutagenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three to four families of nuclear genes encode different isoforms of phosphoenolpyruvate (PEP) carboxylase (PEPC): C4-specific, C3 or etiolated, CAM and root forms. C4 leaf PEPC is encoded by a single gene (ppc) in sorghum and maize, but multiple genes in the C4-dicot Flaveria trinervia. Selective expression of ppc in only C4-mesophyll cells is proposed to be due to nuclear factors, DNA methylation and a distinct gene promoter. Deduced amino acid sequences of C4-PEPC pinpoint the phosphorylatable serine near the N-terminus, C4-specific valine and serine residues near the C-terminus, conserved cysteine, lysine and histidine residues and PEP binding/catalytic sites. During the PEPC reaction, PEP and bicarbonate are first converted into carboxyphosphate and the enolate of pyruvate. Carboxyphosphate decomposes within the active site into Pi and CO2, the latter combining with the enolate to form oxalacetate. Besides carboxylation, PEPC catalyzes a HCO3 --dependent hydrolysis of PEP to yield pyruvate and Pi. Post-translational regulation of PEPC occurs by a phosphorylation/dephosphorylation cascade in vivo and by reversible enzyme oligomerization in vitro. The interrelation between phosphorylation and oligomerization of the enzyme is not clear. PEPC-protein kinase (PEPC-PK), the enzyme responsible for phosphorylation of PEPC, has been studied extensively while only limited information is available on the protein phosphatase 2A capable of dephosphorylating PEPC. The C4 ppc was cloned and expressed in Escherichia coli as well as tobacco. The transformed E. coli produced a functional/phosphorylatable C4 PEPC and the transgenic tobacco plants expressed both C3 and C4 isoforms. Site-directed mutagenesis of ppc indicates the importance of His138, His579 and Arg587 in catalysis and/or substrate-binding by the E. coli enzyme, Ser8 in the regulation of sorghum PEPC. Important areas for further research on C4 PEPC are: mechanism of transduction of light signal during photoactivation of PEPC-PK and PEPC in leaves, extensive use of site-directed mutagenesis to precisely identify other key amino acid residues, changes in quarternary structure of PEPC in vivo, a high-resolution crystal structure, and hormonal regulation of PEPC expression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00029380
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    Electronic Resource
    Stably transformed cell lines from protoplasts of maize endosperm suspension cultures (1994)
    Springer
    Plant cell, tissue and organ culture 37 (1994), S. 39-46 
    Details
    ISSN: 1573-5044
    Keywords: endosperm cell culture ; maize ; protoplast ; transformation ; zeins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts were isolated from Zea mays (L.) A69Y endosperm suspension cultures and transformed by polyethylene glycol mediated DNA uptake with chimaeric gene constructs containing β-glucuronidase (GUS) or neomycin phosphotransferase II (NPTII); GUS-expressing and Kanamycin-resistant cultures were recovered. The transformed cells showed integration of the introduced foreign genes into genomic DNA and maintained their ability to synthesize endosperm-specific reserve proteins (zeins). No deletion or rearrangement of zein genes were observed in transformed cultures. Stable transformation of cultured maize endosperm cells may therefore represent a new methodological approach for the study of the transcriptional regulation of endosperm-expressed genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00048115
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  • 86
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    Efficient plant regeneration through somatic embryogenesis from callus induced on mature rice embryos (Oryza sativa L.) (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 259-264 
    Details
    ISSN: 1573-5044
    Keywords: monocotyledons ; Oryza sativa L. ; plant regeneration ; rice ; somatic embryogenesis ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To obtain a reproducible efficient procedure for regeneration of rice plants through somatic embryogenesis from callus four published methods of callus induction and regeneration were compared. Callus was initiated from mature embryos of the Japonica cultivar Taipei 309 of rice (Oryza sativa L.). The number, mass and morphology of the callus formed on the scutellum were dependent on the medium used. A limited humidity and an optimal aeration of the culture vessels enhanced the frequency of embryogenesis and plant regeneration. A method described by Poonsapaya et al. (1989) was found to be the most efficient and was slightly modified. As a result 98% of the T309 embryos formed callus, of which 63% regenerated into plants. Each callus yielded an average of 6 plants. Plant morphology, fertility and seed set of the regenerants were found to be normal.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00037729
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  • 87
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    Electronic Resource
    Factors that effect leaf regeneration efficiency in apple, and effect of antibiotics in morphogenesis (1994)
    Springer
    Plant cell, tissue and organ culture 37 (1994), S. 257-269 
    Details
    ISSN: 1573-5044
    Keywords: adventitious shoots ; Malus x domestica Borkh. ; tissue culture ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Several factors that affect the frequency of organogenesis in apple leaf explants were examined for the scion cultivars ‘Empire’, ‘Freedom’, ‘Golden Delicious’, ‘Liberty’, ‘McIntosh’, and ‘Mutsu’ and for the rootstocks Malling 7A and Malling 26. The main factors affecting morphogenesis were BA concentration, basal medium, leaf explant origin and maturity, explant orientation, and photosynthetic photon flux. Depending on the genotype, optimal regeneration was obtained using either 22.2 or 31.1 μM BA and the N6 basal medium, with the exception of ‘Golden Delicious’ which regenerated better on MS medium. After 6 weeks, the average number of shoots per segment varied from 5 to 16, and the percentage of regeneration between 70 and 100%, depending on the genotype tested and the maturity of the explant. Regeneration capacity increased dramatically from the tip towards the base of the leaf, and was higher from the middle to the proximal end. Cefotaxime and carbenicillin, two antibiotics commonly used during transformation studies to eliminate Agrobacterium tumefaciens from plant tissue, were tested to determine their effect on morphogenesis. Cefotaxime at a dose of 250 mg 1-1 enhanced regeneration and shoot development, whereas carbenicillin at a dose of 500 mg l-1 induced abundant callus formation and inhibited regeneration. Kanamycin, a widely used selection agent for plant transformation, strongly inhibited regeneration even at very low doses. Schemes for selection and recovery of transgenic apple plants when kanamycin is used as the selection agent are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00042339
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  • 88
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    Micropropagation of thirteen Malus cultivars and rootstocks, and effect of antibiotics on proliferation (1994)
    Springer
    Plant growth regulation 15 (1994), S. 55-67 
    Details
    ISSN: 1573-5087
    Keywords: apple (Malus × domestica Borkh.) ; selection ; tissue culture ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Several factors that affect in vitro establishment, proliferation, and rooting of thirteen Malus cultivars and rootstocks were studied. Apple shoot tips (1.5±0.5 cm in length) were established using ascorbic and citric acids as antioxidants. Four proliferation media containing 1.0 mg 1−1 BA and different concentrations of IBA and GA3 were tested. Proliferation rates varied depending on the genotype and medium used. The highest proliferation rate was obtained for a rootstock that produced 11.6±2.5 shoots (1.5±0.8 cm in length) per tube per month. Rooting was induced with IBA for all the genotypes tested. The optimal IBA concentration was cultivar dependent (between 0.1 and 1.0 mg 1−1 IBA), and lower concentrations were necessary to induce rooting in liquid rather than in solid medium. The effects on shoot-tip proliferation of cefotaxime, carbenicillin and kanamycin, three antibiotics commonly used for transformation studies, were also evaluated. Cefotaxime at 200 mg 1−1 stimulated shoot growth and development, but at 500 mg 1−1 caused abnormal shoot morphology. Carbenicillin at 500 mg 1−1, alone or in combination with cefotaxime at 200 mg 1−1, inhibited proliferation and caused excessive enlargement of the basal leaves, inducing callus formation and release of phenolic compounds in the medium. Kanamycin at 50 mg 1−1 was phytotoxic and caused shoot chlorosis and necrosis. Consideration of the toxicity of these antibiotics is critical when designing transformation schemes for selection and recovery of transgenic apple plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00024677
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  • 89
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    Electronic Resource
    Design and application of antisense oligonucleotides in cell culture,in vivo, and as therapeutic agents (1994)
    Springer
    Cellular and molecular neurobiology 14 (1994), S. 557-568 
    Details
    ISSN: 1573-6830
    Keywords: antisense oligonucleotides ; gene expression ; pharmacology ; drug design ; cell cultures ; brain research
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. Synthetic oligonucleotides can inhibit the expression of a gene in a sequence specific manner on the transcriptional and translational level. These molecules are usually referred to as antisense oligonucleotides. 2. Antisense mediated inhibition of gene expression is a valuable tool to analyze the function of a genein vivo and can also be used for therapeutic gene suppression. 3. A number of factors such as the mode of action, specificity, chemistry, and pharmacology must be carefully considered for the design and successful application of antisense oligonucleotides. 4. Assay systems and controls must be chosen as to assure that the observed biological effects of antisense oligonucleotides do in fact reflect the result of a specific gene inhibition. 5. This article critically discusses these factors in view of the literature and our own experience with a wide range of cell types and animal models, targeting different genes. The emphasis is on the use of phosphorothioate oligodeoxynucleotides in cell cultures,in vivo, and as potential drugs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02088837
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  • 90
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    Nitrostyrene Derivatives of Adenosine 5′-Glutarates Modified with an Alkyl Spacer and their inhibitory activity on epidermal growth factor receptor protein tyrosine kinase (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994), S. 59-69 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: β-Nitrostyrene derivatives of adenosine 5′-glutarates are potent and selective bisubstrate-type inhibitors of the epidermal growth factor receptor protein tyrosine kinase (EGF-R PTK). In an attempt to improve the inhibitory activity, this type of compounds was modified with alkyl spacers of varying length between the nitrostyrene and the glutaryl units. The spacers consisted of 1, 3, 4, and 5 atoms to give compounds of the benzyl, oxyethyl, oxypropyl, and oxybutyl series, respectively (Schemes 1 and 2). Adenosine 5′-esters were prepared in the benzyl and oxypropyl series only. Compared to the compounds in the parent series without spacer (IC50 = 0.7-12 μM), most of the modified compounds inhibited the EGF-R PTK only marginally or were inactive (IC50 ≥ 100 μM). The only exceptions were the free acids 19 and 20 with IC50 values of ca. 5 μM. It is noteworthy that esterification of these two hydrogen glutarates with either MeOH or adenosine yielded inactive compounds, which is in contrast to the corresponding substances without spacers.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770109
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    Synthese und Eigenschaften der Furo- und Thieno-Analogen von PQQ-TriesterTeil einer Präsentation am ‘2nd International Symposium on PQQ and Quinoproteins’, 19.-22. Nov. 1991, Yamaguchi, Japan. (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994), S. 100-110 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Synthesis and Characterization of the Furo and Thieno Analogues of the Triester of PQQWe report here the synthesis and properties of the furo and thieno analogues of 4,5-dihydro-4,5-dioxo-1H-pyrrolo[2,3-f]quinoline-2,7,9- tricarboxylic acid ( = PQQ), i.e. the furo- and thieno[2,3-f]quinoline-4,5-quinone (FQQ and TQQ, resp.) derivatives B and C, obtained as triesters. The triester of PQQ derivative A is much more stable than the triesters of B or C, and only the triester of A shows strong activity in nonenzymatic catalytic oxidations.
    Additional Material: 1 Tab.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770113
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  • 92
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    Computational Approach to the Receptor-Bound Conformation of Cyclosporin A (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994), S. 171-181 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The conversion of the conformation of cyclosporin A (CsA) observed in CHCl3 to the receptor-bound state is investigated by two molecular-mechanics methods, template forcing and dynamic forcing. The conformations of CsA in CHCl3 and complexed with LiCl in THF as determined by NMR are used as starting structures. The transition starting from the CsA/CHCl3-derived conformation is hindered by steric interactions of two side chains (MeBmt1 and Val5). While starting with the CsA/LiCl-derived conformation, the conversion is facile. It is illustrated that these calculations, which are of artificial character, using only the starting and final structures of the observed conformational transition during the receptor-binding event, allow an insight into the interactions between the substrates and receptor in terms of an induced fit.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770120
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  • 93
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    Electrochemical Study of Phase-Transfer Catalysis Reactions: The Williamson ether synthesis (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994), S. 231-242 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The transfer properties of the ionic species involved in the Williamson ether synthesis by phase-transfer catalysis were investigated using electrochemical techniques developed for the study of polarised liquid/liquid interfaces. This approach allows the measurement of the apparent partition coefficients of the transferring species. From these data, it is proposed that the role of the phase-transfer catalyst salt in the reaction mechanism is to establish a Galvani distribution potential difference between the two phases which in turn acts as the driving force for transferring the reactive aqueous ions to the organic phase.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770125
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  • 94
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    Synthesis of (R)- and (S)-5-(Hydroxymethyl)-3-isopropyloxazolidin-2-one, intermediates in the preparation of optically active β-blockers (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994), S. 252-256 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The (R)- and (S)-5-(hydroxymethyl)-3-isopropyloxazolidin-2-ones, ((R)- and (S)-2, resp.), pivotal intermediates in the preparation of optically active β-blockers, were synthesized using (R,E)-2-hydroxypent-3-enenitrile (1) as the chiral starting material. In the synthesis of (R)-2, a known cyclization/inversion step was applied.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770127
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  • 95
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    Calorimetric Measurements of the Complexation of Cyclosporin A, Ascomycin, Fujimycin, and Rapamycin with Lithium Chloride and with an Immunophilin (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994), S. 291-305 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Solutions (2 ml) of small linear and cyclic peptides (4-11), of a peptolide containing nine amino acids and a lactate moiety (12), of the cyclic undecapeptide cyclosporin A (CS, 1), and of the macrolides ascomycin, fujimycin, and rapamycin (13-15) in THF were added to excess LiCl, LiBr, or LiClO4 (up to 3000 equiv. in 40 ml THF) in a calorimeter (calorimetric titration). The enthalpies of interaction measured are in the range of ΔH = -8 to -37 kcal/mol. A similar experiment was carried out with one of the binding proteins of cyclosporin, the human cyclophilin A, to give the thermodynamic parameters for the complexation ΔH = -16, ΔG° = -10 kcal/mol, and ΔS° = -20 cal/mol·deg. at 25° which corresponds to an equilibrium constant K = 2·107 l/mol, in good agreement with the result of independent measurements using different methods. NMR Measurements of the macrolides in (D8)THF containing LiCl show strong down-field shifts of signals of the H-atoms next to C=O and C-OH groups in these molecules.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770129
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  • 96
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    NMR of Terminal Oxygen. Part 13Part 12: [1].. 17O-NMR spectra of C-nitroso compounds, thionitrites and NO+ Ion: Resonance effects in O=N - X compounds and correlation with CD spectra (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994), S. 306-316 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The 17O-NMR signals of four true C-nitroso compounds 1-4 appear at particularly low field (1550-1265 ppm), whereas the dimers (azodioxy type) resonate at ca. 400 ppm and the ‘isonitroso compounds’ ( = quinone-oximes; 5 and 6) at ca. 250 ppm. S-Nitroso compounds ( = thionitrites; 8 and 9) show shift values of ca. 1300 ppm, not far from C - NO; the NO+ ion is much stronger shielded (474 ppm). The results, together with those for higher-shielded nitroso compounds X - NO (X = RO, R2N, Cl, O-) are discussed in terms of (a) resonance stabilization through n-donation from X(π-bond order, approximated by the known barriers of rotation around the X - N bond) and of (b) electronic excitation energies ΔE. The latter are approximated by long-wave (symmetry-forbidden) UV/VIS absorptions and confirmed, where available, by the maxima of the curves of circular dichroism (CD); the CD curve of thionitrite 9 has been measured. It is found that the δ(17O) values of X - NO depend both on bond order and on ΔE, which could not be separated. The higher shielding of NO+ compared with X - N=O is explained on the basis of anisotropy effects, which differ between sp and sp2 systems.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770130
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  • 97
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    Electronic Resource
    Masthead (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994) 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770101
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  • 98
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    Enantioselective synthesis of (-)-conduramine C1 and aminobromocyclitol derivatives‘Naked Sugars’ as Synthetic Intermediates, Part XXV. Part XXIV, see [1a]; Part XXIII, see [1b]. (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994), S. 1-9 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: (1S,2R,6R,7R)-4-Phenyl-3,10-dioxa-5-azatricyclo[5.2.1.02,6]dec-4-en-9-one ((+)-5) obtained in 6 steps from the Diels-Alder adduct of furan to 1-cyanovinyl (1S)-camphanate ((+)-3) was reduced to the corresponding endo-alcohol (-)-6 the treatment of which with HBr/AcOH provided (-)-(3aS,4S,6R,7S,7aR)-4β-bromo-3aβ,4,5,6,7,7aβ-hexahydro-2-phenyl-1,3-benzoxazole-6β,7α-diyl diacetate ((-)-17). Elimination of HBr with 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and acidic hydrolysis furnished (-)-(1R,2S,3R,4R)-4-aminocyclohex-5-ene-1,2,3-triol ( = (-)-conduramine C1;(-)-1).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770103
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  • 99
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    Nucleotides Part XLPart XXXIX: [1].. Synthesis and Characterization of Modified 2′-5′ Adenylate Trimers-Potential Antiviral Agents (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994), S. 10-22 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: 2′-5′ Adenylate trimers 41-44 carrying the (tert-butyl)dimethylsilyl (tbds) group at the 3′-OH position of various sugar moieties were synthesized via the phosphoramidite method. The use of the (tert-butyloxy)carbonyl (boc) and 2-(4-nitrophenyl)ethylsulfonyl (npes) groups for 2′-OH protection in neighbourhood to the 3′-O-tbds residue was compared during the synthesis of the target trimers. For other functional positions, the use of the 2-(4-nitrophenyl)ethyl (npe) and 2-(4-nitrophenyl)ethoxycarbonyl (npeoc) blocking groups were favoured.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770104
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  • 100
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    Metal complexes of macrocyclic ligands. Part XXXVIIPart XXXVI: [1].. Synthesis of Heteroditopic Bis-macrocycles and Their Potential for Preparing Heterobinuclear Metal Complexes (1994)
    New York, NY : Wiley-Blackwell
    Helvetica Chimica Acta 77 (1994), S. 23-35 
    Details
    ISSN: 0018-019X
    Keywords: Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A new and generally applicable synthetic path for the preparation of heteroditopic bis-macrocycles using tri-N-protected tetraazacycloalkanes as building blocks and bromoacetyl bromide as bridging reagent is described. In the first step, bromoacetyl bromide is used as acylating agent for one of the macrocycles, whereas in the second step it is used as alkylating agent for the second macrocycle, thus giving protected bis-macrocyclic amides (e.g. 6). After reduction of the amide moiety and deprotection, bis-azamacrocycles with an ethylene bridge are obtained (e.g. 8). The corresponding homoditopic bis-macrocycles 16 and 17 are also prepared for comparison purpose. Spectrophotometric studies indicate that bis-macrocycle 8, which consists of a 12- and a 14-membered ring, binds two metal ions with equal affinity, whereas compound 13, in which an unsubstituted (cyclam) and a trimethyl-substituted tetraazacyclotetradecane unit (Me3cyclam) are bridged, shows selective metal-ion binding. The first metal ion is always incorporated into the cyclam unit, whereas the second one binds to the Me3cyclam macrocycle. Thus, by sequential addition of two different metal ions, heterobinuclear complexes can easily be prepared. The electrochemistry of the binuclear Ni2+ complexes, studied by CV and DPV, as well as the EPR spectra of the binuclear Cu2+ complexes clearly indicate metal-metal interactions.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/hlca.19940770105
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