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  • 1985-1989  (78)
  • 1955-1959  (17)
  • 1890-1899
  • 1820-1829
  • somaclonal variation
  • transformation
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 16 (1989), S. 369-372 
    ISSN: 1432-0983
    Keywords: Bialaphos ; Neurospora crassa ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Conidia of Neurospora crassa are sensitive to the herbicide bialaphos at concentrations of 160 mg/1 in Westergaard's or Fries' minimal media. Plasmid pJA4 was constructed by inserting a truncated bar gene from Streptomyces hygroscopicus fused to the his-3 promoter from N. crassa into pUC19. The bar gene in plasmid pJA4 confers resistance to bialaphos when transformants are selected on a medium containing bialaphos. The bar gene can be used as an additional dominant selectable marker for transformation of fungi.
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  • 2
    ISSN: 1436-6215
    Keywords: γ-Tocopherol ; Transformation ; α-Tocopherol ; Ratten ; Generationsversuch ; γ-tocopherol ; transformation ; α-tocopherol ; rats ; generation-experiment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Summary The biosynthesis of α-tocopherol, the most effective vitamer among the vitamin E-group, is found only in higher plants and microorganisms. Due to the lack of the shikimate pathway, animals are not able to synthesize α-tocopherol. Also not found is a whole enterai synthesis; only the conversion of dimethyletocol to trimethyletocol seems to be possible. Using four generations of rats, we sought to determine: Is a transformation of γ-tocopherol to α-tocopherol in the animal body possible? Are there any differences in the transformation rates in organs, tissues, or in the entire body along the generations? Does gut flora play any role in the conversion of γ-tocopherol? Is it possible to increase the efficiency of the transformation by supplying additional CH3-groups? Wistar rats were fed a semisynthetic basal diet, supplemented with 78.8 mg DL-γ-tocopherol/kg in the first three generations (F1–F3). In the fourth generation (F4), some of the animals were fed a vitamin E-free diet and γ-tocopherol (approx. 1.5 mg on alternate days) was injected s.c. Two other groups of animals received the basal diet containing additional methionine (0.25 %) or choline (0.45 %), as well as γ-tocopherol (as in F1–F3). α- and γ-tocopherol were analyzed by HPTLC in the whole body and in serum, liver, heart, lung, gut, gonads, and feces. The ratio of α-/γ-tocopherol (μg/μg) as transformation rate and vitamin E-biopotency (μg α-tocopherol equivalents/g) were calculated. Growth and fertility were normal until the fourth generation; no abnormal developments could be recognized. α-tocopherol was found in the whole-body as well as in all tissues and organs. In the whole-body, vitamin E-biopotency decreased 25–70 % in F2 and F3. On the other hand, the increase of the transformation rate of γ- to α-tocopherol amounted to 23 % (F2) and 168 % (F3). Highest conversion rates were found in F2 and F3 for feces, followed by gonads and lungs; the lowest rates were found for serum and liver. Due to the s.c.-injection of γ-tocopherol, feces showed a four-times lower transformation rate in F4 than in F3. There was an increase in heart, gut, lung and serum for both transformation rate and vitamin E-biopotency. These parameters could be improved also by the additional supplements of methionine and choline. Both methyl-group-donators revealed nearly the same positive effect. The results show that the animal organism can adapt to γ-tocopherol supply over generations. γ-tocopherol seems to be a direct precursor for the α-tocopherol synthesis. The methylation of γ-tocopherol in the organs and tissues occurs, presumably, according to their specific α-tocopherol requirement.
    Notes: Zusammenfassung Die Biosynthese des α-Tocopherols, des wirksamsten Vitamins innerhalb der Vitamin-E-Gruppe, ist beschränkt auf höhere Pflanzen und Mikroorganismen. Wegen des Fehlens des Shikimatweges vermag der tierische Organismus das α-Tocopherol nicht zu bilden. Auch eine vollständige enterale Synthese ist nicht bekannt. Es wird angenommen, daß die Umwandlung im Tierkörper von Dimethyltocol zum Trimethyltocol möglich sei. In einem Experiment an Ratten über vier Generationen wurde folgenden Fragen nachgegangen: Findet eine Umwandlung von γ- zu α-Tocopherol statt? Ändert sich die Effizienz der Transformation auf Gewebs- und Organebene bzw. im gesamten Körper über die Generationen? Welche Rolle spielt die Darmflora? Kann die Effizienz der Transformation durch zusätzliche Gaben an CH3-Gruppen verbessert werden? Über vier Generationen erhielten Wistarratten eine halbsynthetische Grunddiät mit 78,8 mg DL-γ-Tocopherol/kg (F1–F3). In F4 erhielt ein Teil der Tiere die tocopherolfreie Grunddiät, γ-Tocopherol (ca. 1,5 mg, alle zwei Tage) wurde den Tieren subkutan verabreicht. Weitere zwei Kollektive bekamen mit dem Grundfutter γ-Tocopherol (wie in F1–F3) und zusätzlich Methionin (0,24 %) bzw. Cholin (0,45 %) oral verabreicht. In einer Ganzkörperanalyse in F1–F3 und in Serum, Erythrozyten, Leber, Herz, Lunge, Darm, Gonaden und Kot wurden α- und γ-Tocopherol mittels HPTLC bestimmt. Das Verhältnis α-γ-Tocopherol (μg/μg) und die Vitamin-E-Wirksamkeit (μg α-Tocopheroläquivalente/ml bzw. g FS od. g TS) wurden errechnet. Bis zur 4. Filialgeneration waren Wachstum und Fortpflanzungsfähigkeit normal; keine äußeren oder anatomischen Abnormitäten wurden beobachtet. Im Gesamtkörper und in Geweben und Organen der Generationen F1–F4 wurde α-Tocopherol gefunden. Gemessen an den Ergebnissen der Ganzkörperanalyse nahm die Vitamin-E-Wirksamkeit in F2 um 25 % und in F3 um 70 % ab. Die Effizienz der γ-Tocopheroltransformation stieg dagegen um 23 % in F2 und 168 % in F3. Höchste Transformationsraten wurden in F2 und F3 für Kot, gefolgt von Gonaden und Lunge, festgestellt, die niedrigsten für Serum und Leber. Durch die subkutane γ-Tocopherolapplikation war die Transformationsrate im Kot in F4 um Faktor 4 schlechter als in F3. Die Effizienz der Transformation und die Vitamin-E-Wirksamkeit nahmen in Herz, Dickdarm, Lunge und Serum zu. Ebenso besser fielen die Werte für diese Parameter unter der Mehrzufuhr an Methionin und Cholin, wobei sich mit beiden Methylgruppendonatoren die gleiche positive Wirkung erzielen ließ. Die Ergebnisse zeigten, daß sich der Körper an eine γ-Tocopherolzufuhr über Generationen adaptieren kann. γ-Tocopherol dient auch im tierischen Organismus als unmittelbare Vorstufe der α-Tocopherolsynthese. Dieser Syntheseschritt erfolgt wahrscheinlich über eine Transmethylierungsreaktion in den verschiedenen Geweben und Organen gemäß ihrem spezifischen Bedarf an α-Tocopherol.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Methods in cell science 12 (1989), S. 127-133 
    ISSN: 1573-0603
    Keywords: polyethylene glycol ; transformation ; protoplasts ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Polyethylene glycol can be used to induce DNA uptake into plant protoplasts. Procedures for isolation, culture and transformation ofN. tabacum protoplasts are described and can be adapted for other dicot and monocot species. Criteria for proof of transformation are discussed.
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  • 4
    ISSN: 1573-5028
    Keywords: Agrobacterium rhizogenes ; border sequence ; T-DNA ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A Charon 4A phage library, containing insert DNA isolated from a morning glory (Convolvulus arvensis) plant genetically transformed by Ri T-DNA from Agrobacterium rhizogenes strain A4, was used to isolate a lambda clone that contains part of the Ri TL-DNA and the complete TR-DNA. The two Ri T-DNAs were recovered adjacent to each other in a tail-to-tail configuration (i.e. with the TR-DNA inverted with respect to the TL-DNA). Comparison of nucleotide sequences from this lambda clone with the corresponding sequences from the Ri plasmid allowed us to determine the location of the T-DNA/plant junction for the right end of the TL-DNA and the left and right ends of the TR-DNA. We located, near each of these borders, a 24 bp sequence that is similar to the 24 bp consensus sequence found near the pTi T-DNA extremities. In addition, sequences similar to the “core” overdrive sequence from pTi are located near each right border. Hybridization and nucleotide sequence analysis of the DNA adjacent to the TL/TR junction shows that no plant DNA is located between the TL and TR-DNAs and suggests that the plant DNA adjacent to the end of the TR-DNA may have been rearranged during the integration into the plant genome.
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  • 5
    ISSN: 1573-4943
    Keywords: conformational energy ; amino acid substitution ; P21 protein ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Substitutions of amino acids for Gly 12 or Gly 13 in theras oncogene-encoded P21 proteins have been demonstrated to produce unique structural changes in these proteins that correlate with their ability to produce cell transformation. For example, the P21 proteins with Arg 12 or Val 13 are both known to be actively transforming. Recent site-specific mutagenesis experiments on the transforming Arg 12 protein have found that the substitution of Val for Gly 10 has no effect on transforming activity whereas the substitution of Val for Gly 13 led to a loss of transforming activity. In this study, we examine the structural effects of these substitutions on the amino terminal hydrophobic decapeptide (Leu 6-Gly 15) of P21 using conformational energy analysis. The results show that the transforming proteins with Gly 10 and Arg 12 or Val 10 and Arg 12 can both adopt the putative malignancy-causing conformation, whereas, for the nontransforming protein with Arg 12 and Val 13, this conformation is energetically disallowed. These results further support the theory that due to structural changes the transforming P21 proteins are unable to bind to some regulatory cellular element which may be the recently identified binding protein responsible for the induction of increased GTPase activity in normal P21 compared with transforming mutants.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 13 (1989), S. 151-161 
    ISSN: 1573-5028
    Keywords: β-glucuronidase (gusA) gene ; maize ; protoplasts ; stable co-transformation ; transformation ; Zea mays L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An efficient co-transformation protocol using polyethylene glycol was developed for Zea mays L. (cv. A188 × BMS) protoplasts isolated from suspension culture cells. Co-transformation was accomplished by using plasmid constructions containing β-glucuronidase (gusA) or neomycin phosphotransferase (neo) gene coding sequences; both were under control of the CaMV 35S promoter. Protoplast culture and transformation conditions were optimized to assure efficient recovery of transformed cells. The overall efficiency of transformation was 1 × 10−4 (calculated per viable protoplast plated). Among kanamycin-resistant lines, 50% showed a high level of GUS activity (above one unit). Southern blot hybridization confirmed the presence of numerous gusA and neo coding sequences in the maize genome. In two analyzed lines, integrated sequences appeared to be organized in tandem head-to-tail repeats. Results also indicated that the integrated sequences were partially methylated.
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  • 7
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; adventitious shoot regeneration ; transformation ; homozygous potato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transformed potato (Solanum tuberosum) plants were obtained from homozygous diploid potato by using a transformation procedure in combination with an adventitious shoot regeneration method. Leaf and stem explants were inoculated with an Agrobacterium tumefaciens strain which contained a binary vector (pVU 1011) carrying the neomycin phosphotransferase gene. Shoot regeneration most effectively on stem explants, occurred within six weeks directly from the explants without introducing a callus phase. A strong seasonal influence on transformation efficiencies was observed. Analysis of a number of randomly selected regenerated shoots for their ability to root and form shoots on kanamycin-containing medium shows that over 90% of the regenerated shoots obtained are transformed. In a number of shoots transformation was confirmed by a test for the presence and expression of the NPT-II gene.
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  • 8
    ISSN: 1573-5028
    Keywords: herbicide ; mutant ; photosystem II ; psbA gene ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mutations conferring herbicide resistance in 3 mutant strains of the cyanobacterium Synechocystis 6714 have been characterized by gene cloning and sequencing. The mutants display very different phenotypes: DCMU-IIA is DCMU-resistant and atrazine-resistant, DCMU-IIB is DCMU-resistant and atrazine-sensitive, and Az-V is DCMU-sensitive, atrazine-resistant and presents particular photoinhibition properties. These mutants were originally obtained either by one-step selection (DCMU-IIA) or by two-step selection (DCMU-IIB and Az-V). psbA copies carrying herbicide resistance have been identified by transformation experiments as psbAI in all cases. Sequences of the psbAI copy of each mutant have been compared to the wild-type sequence. In the single mutant DCMU-IIA, a point mutation at codon 264 (Ser→Ala) results in resistance to both DCMU and atrazine. In the double mutants DCMU-IIB and Az-V, two point mutations were found. DCMU-IIB was derived from DCMU-IIA and had acquired a second mutation at codon 255 (Phe→Leu) resulting in a slight increase in DCMU resistance and complete abolition of atrazine resistance. Az-V contains two changes at codons 211 (Phe→Ser) and 251 (Ala→Val) resulting in high atrazine resistance but only slight DCMU resistance.
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  • 9
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; C4 photosynthesis ; Flaveria ; regeneration ; ribulose bisphosphate carboxylase/oxygenase ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the successful transformation, via Agrobacterium tumefaciens infection, and regeneration of two species of the genus Flaveria: F. brownii and F. palmeri. We document the expression of a C3 plant gene, an abundantly expressed ribulose 1,5-bisphosphate carboxylase/oxygenase small subunit gene isolated from petunia, in these C4 plants. The organ-specific expression of this petunia gene in Flaveria brownii is qualitatively identical to its endogenous pattern of expression.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 42 (1989), S. 155-162 
    ISSN: 1573-5060
    Keywords: Rubus laciniatus ; thornless blackberry ; brambles ; tissue culture ; periclinal chimera ; in vitro ; ex vitro ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary ‘Thornless Evergreen’ (TE) blackberry (Rubus laciniatus Willd.) is a periclinal chimera in which the epidermis has mutated to thornlessness (designated ‘t’), while the internal portions of the plant remain genetically thorny (tTT). Over 300 ex vitro plants (from shoot tip cultures of TE) were established in the field and observed over two fruiting seasons for various vegetative traits and fruiting characteristics. Both parental types and variants were included in the field plantings. Plants were evaluated for growth habit, vigor, flowering, and fruiting traits. Continuous variation for growth habit, vigor, flower number, and fertility was observed. Some of the variation could be explained in terms of chimeral segregation (pure thornless vs. chimeral types). Tissue culture-induced (somaclonal) variation was assumed to be the source of most of the other variation.
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  • 11
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 40 (1989), S. 75-81 
    ISSN: 1573-5060
    Keywords: Brassica juncea ; Indian mustard ; plant generation ; somaclones ; yield ; oil content ; dwarf type ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Multiple shoot formation in cotyledonary callus of Indian mustard (Brassica juncea cv. Prakash) was induced on modified MS media supplemented with high cytokinin (kinetin or zeatin) and low IAA concentrations. Complete plants were obtained on prolonged incubation of shoots on the same medium. 6-Benzyladenine alone or in combination with IAA or NAA did not support plantlet regeneration. A total of 71 plants were transferred to greenhouse. The seed, however, could be collected from 37 plants only. The seed was sown in the field to evaluate the material for somaclonal variation in R1 generation. Data were recorded for yield, plant height, number of primary branches, siliqua number, 1,000 seed weight and oil content. Somaclonal lines showed tremendous amount of variation for all the characters studied. A number of plants in this generation showed significantly higher yield and/or other improved agriculturally important characteristics as compared to the control. A line with dwarf plant type was also identified. A number of plants were selected from this generation and carried forward to R2 generation. Most of these lines bred true in R2 generation. The material seems to be very promising for future breeding programmes.
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 19 (1989), S. 159-165 
    ISSN: 1573-5044
    Keywords: cultivars ; roots ; shoots ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four tronchuda (Brassica oleracea var. tronchuda Bailey) cultivars were tested for their ability to regenerate in vitro on Murashige & Skoog (MS) medium supplemented with 3 different combinations of α-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP). Explants were either axillary bud-free whole cotyledons or hypocotyls from 7-day-old darkgrown seedlings. The ability to regenerate varied by cultivars, explants and the concentration of growth regulators. Hypocotyl explants of all 4 cultivars, and cotyledon explants of 2 cultivars, developed plantlets within 4 weeks. Hypocotyl explants produced more shoots than cotyledons. Cotyledon explants produced more roots than hypocotyls. Best shoot regeneration was on MS medium supplemented with 2 mgl-1 BAP and 0.1 mgl-1 NAA. ‘Portuguesa’ produced the most shoots. Some regenerants varied in leaf shape and phyllotaxy.
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Methods in cell science 12 (1989), S. 135-138 
    ISSN: 1573-0603
    Keywords: transformation ; Agrobacterium tumefaciens ; vectors ; cocultivation ; leaf disc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Agrobacterium tumefaciens naturally transfers DNA into plant cells and is clearly one of the most effective methods of directed DNA transfer presently available. Two kinds of vectors are commonly used. Cointegrative vectors have the foreign genes incorporated directly into the Ti plasmid. Binary vectors carry two plasmids; the main Ti plasmid where most of the T-DNA has been removed, and a second plasmid containing the foreign genes between the usual border sequences. The vir genes on the main plasmid function to mobilize the foreign genes into a plant cell. Most plant transformation methods follow the procedure of cocultivating wounded tissue with vir-gene-induced bacteria. The cocultivation step is followed by transfer to a selective medium containing antibiotics to kill the bacterium and to allow only growth of transformed tissue. Several selectable markers are available that include resistance to antibiotics, herbicides, or drugs. In addition, several scorable markers such as the bacterial glucuronidase, chloramphenicol acetyl transferase, and the Agrobacterium opine genes are used to verify transformation. Southern blotting and inheritance of transferred genes are ultimately used to demonstrate stable transformation.
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  • 14
    ISSN: 1573-5044
    Keywords: Oryza sativa L. ; somaclonal variation ; grain and quality characters ; protein content
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Variations in some grain and quality characters in progenies of regenerated rice plants were studied. Grain length and weight decreased significantly, yet gel consistency increased. Variations in these quantitative characters of all cultivars studied were consistent, showing the tendencies of the variations. Grain protein contents of the somaclones were higher in one cultivar. Variability of most traits was increased by combining low-dosage radiation and tissue culture.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 18 (1989), S. 281-296 
    ISSN: 1573-5044
    Keywords: maize cells ; transformation ; kanamycin electroporation ; nurse cells ; hygromycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two chimaeric genes, containing the promoter of the 35S gene of cauliflower mosaic virus coupled to neomycin phosphotransferase (35S-NPT-NOS) or to hygromycin phosphotransferase (35S-HPT-NOS) have been stably transferred to maize (Zea mays cv. Black Mexican sweet) cells by electroporation. Transformation frequencies of 7.6×10-4 and 8×10-4, respectively, (based on the number of surviving cells that divided) were obtained with four pulses of 1 ms duration using 400 V capacitive discharge. Cells transformed to kanamycin-resistance and hygromycin-resistance subsequently multiplied to form callus. Southern blot analysis demonstrated the integration of the selectable marker genes, neomycin or hygromycin phosphotransferase, with single or multiple copy numbers. The blots from DNA of hygromycin-resistant calli also suggested the formation of plasmid concatemers.
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  • 16
    ISSN: 1573-0778
    Keywords: bronchus ; cell culture ; cytology ; morphometry ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Malignant A-549 lung carcinoma and adenovirus-12 SV40 hybrid virus transformed non-tumorigenic human bronchial epithelial cells (BEAS-2B) were objectively discriminated from normal bronchial epithelial (BE) cells on the basis of Papanicolaou stained nuclear features (e.g. shape, chromatin texture, hyperchromasia) and nucleolar morphology (e.g. number per cell, irregular contours). Morphometric analysis indicated that significant differences in cellular morphology existed between BE, BEAS-2B, and A-549 cells. Similar analyses of transformed, tumorigenic cell lines demonstrated that nuclear features (i.e., chromatin texture, clearing of parachromatin, hyperchromasia, variation in thickness of the nuclear envelope, sharp indentations in the nuclear envelope), and nucleolar features (i.e., degree of roundness, presence of angular projections, number per cell) discriminated chemically and virally transformed cells from spontaneously transformed cells. Nuclear and nucleolar features were correlated with the growth rate of tumorigenic cell lines. These analytical approaches will be helpful in studies of the effects of various factors (e.g. vitamin A, phorbol ester, oncogene transfection) on cellular proliferation and/or differentiation.
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  • 17
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Applied Organometallic Chemistry 3 (1989), S. 203-209 
    ISSN: 0268-2605
    Keywords: Phosphorus-carbon ; organisms ; formation ; transformation ; ciliatine ; bialaphos ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Additional Material: 1 Tab.
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  • 18
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 10 (1989), S. 189-197 
    ISSN: 0192-253X
    Keywords: Drosophila ; transformation ; Glue gene ; Gene cluster ; Gene regulation ; Sgs-3 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We reviewed studies on the developmental regulation of the 68C glue gene cluster of Drosophila melanogaster. Extensive transformation analyses of Sgs-3 have shown that four regions necessary for normal expression can be distinguished. The first ( + 10 to -50) contains the transcription start site and TATA motif. This region can be replaced functionally by corresponding sequences from the hsp70 gene, but it is sensitive to point mutations in the TATA sequence. The second region (-50 to -98) contains more than one upstream sequence that, in combination with the other elements, leads to stage and tissue-specific expression. The third region (centered at -600) contains an element that enhances transcript levels some 20-fold. The final region (between -1.65 and -2.35 kb) contains elements having modest (twofold to threefold) effects on expression, one of which is contained in the coding sequences of Sgs-7, a second member of the cluster.
    Additional Material: 4 Ill.
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  • 19
    ISSN: 1573-4943
    Keywords: conformational energy ; amino acid substitution ; position 13 ; P21 protein ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The effect of the substitution of Arg for Gly 13 on the structure of the transforming region decapeptide (Leu 6-Gly 15) of the ras oncogene encoded P21 protein has been investigated using conformational energy analysis. A human malignancy has been identified that contains a ras gene with a single mutation in the thirteenth codon such that the encoded protein would have Arg substituted for Gly at this position, and transfection of cells in culture with this gene results in malignant transformation. Conformational analysis demonstrates that the Arg 13 decapeptide adopts a conformation identical to that for other peptides with substitutions at position 13 (Asp 13, Val 13) from transforming proteins that is distinctively different from that for peptides (Gly 13, Ser 13) from normal, nontransforming proteins. This is found to be an indirect effect resulting from changes in the conformation of Gly 12 produced by substitutions at position 13. These results are consistent with recent analysis of crystallographic data of proteins on conformational preferences for glycine in tripeptide sequences.
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  • 20
    Electronic Resource
    Electronic Resource
    Springer
    Bioscience reports 8 (1988), S. 519-529 
    ISSN: 1573-4935
    Keywords: cell division ; malignancy ; transformation ; nutrient uptake ; resting state
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The problem of regulation of cell division is essentially a problem of understanding regulation of transition from the resting state of a cell to the dividing state and vice versa. In malignancy the ability to revert back to a normal resting state is impaired. A model is presented which attempts to explain the control of the above transitions through control of uptake of essential nutrients by a transport-inhibitory protein. Experimental evidence in favour of the model is given.
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  • 21
    ISSN: 1573-5028
    Keywords: Agrobacterium ; gene expression ; legumin (Pisum) ; Nicotiana ; seed storage protein ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A 3.4-kilobase genomic DNA fragment from Pisum sativum L. containing the LegA gene, which encodes a major legumin storage protein, was transferred to Nicotiana plumbaginifolia using an Agrobacterium tumefaciens strain containing the Bin 19 binary vector system. Northern hybridisation analysis of legA-transformed plants demonstrated that legumin-specific RNA was present in developing seeds but not in developing leaves. Legumin protein was immunologically detected in the mature seeds of legA-transformed plants, and was present as the correct-size protein composed of disulphide-bonded polypeptides. It is concluded that the transferred pea genomic fragment contains all the information necessary for seed-specific expression of the legA gene, and for correct processing of the primary transcript and the precursor legumin protein.
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  • 22
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 11 (1988), S. 551-559 
    ISSN: 1573-5028
    Keywords: Agrobacterium rhizogenes ; Agrobacterium tumefaciens ; Linum usitatissimum ; plant regeneration ; Ri plasmid ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Regeneration of flax (Linum usitatissimum) following transformation by either Agrobacterium tumefaciens carrying a disarmed Ti-plasmid vector, or Agrobacterium rhizogenes carrying an unmodified Ri plasmid, was examined. Hypocotyl and cotyledon explants inoculated with A. tumefaciens formed transformed callus, but did not regenerate transformed shoots either directly or via callus. However, cotyledon explants inoculated with A. rhizogenes formed transformed roots which did regenerate transformed shoots. Ri T-DNA encoded opines were detected in the transformed plantlets and Southern hybridization analysis confirmed the presence of T-DNA from the Ri plasmid in their DNA. Transformed plantlets had curled leaves, short internodes and some had a more developed root system characterized by plagiotropic behaviour.
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  • 23
    ISSN: 1573-5028
    Keywords: plants ; antisense ; suppression ; nopaline synthase ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the successful suppression of nopaline synthase (EC 1.5.1.19) enzymatic activity in the leaves of tobacco plants via the overproduction of RNAs complementary to the nopaline synthase (nos) mRNA. Several different regions of the nos gene were fused, in antisense orientation, to the promoter from a strongly expressed petunia chlorophyll a/b-binding protein gene. These constructions were directly introduced into a tobacco line which contained a single copy of the wild-type nos gene and transgenic plants were regenerated. The degree of nopaline synthase suppression in the leaves of the double transformants ranged up to 85% and was dependent on the particular region of the nos gene present in the antisense RNA. The most effective nos antisense sequences were derived from the 3′ half of the nos gene transcript. In addition, we report a new sensitive method for the detection and quantitation of nopaline synthase activity in crude plant extracts.
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  • 24
    ISSN: 1860-0980
    Keywords: homogeneity analysis ; correspondence analysis ; optimal scaling ; transformation ; alternating least squares ; canonical correlation analysis ; principal component analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Psychology
    Notes: Abstract Homogeneity analysis, or multiple correspondence analysis, is usually applied tok separate variables. In this paper we apply it to sets of variables by using sums within sets. The resulting technique is called OVERALS. It uses the notion of optimal scaling, with transformations that can be multiple or single. The single transformations consist of three types: nominal, ordinal, and numerical. The corresponding OVERALS computer program minimizes a least squares loss function by using an alternating least squares algorithm. Many existing linear and nonlinear multivariate analysis techniques are shown to be special cases of OVERALS. An application to data from an epidemiological survey is presented.
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  • 25
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    Springer
    Psychometrika 53 (1988), S. 437-454 
    ISSN: 1860-0980
    Keywords: multivariate analysis ; optimal scaling ; correspondence analysis ; structural models ; simultaneous equations ; factor analysis ; LISREL ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Psychology
    Notes: Abstract We study the class of multivariate distributions in which all bivariate regressions can be linearized by separate transformation of each of the variables. This class seems more realistic than the multivariate normal or the elliptical distributions, and at the same time its study allows us to combine the results from multivariate analysis with optimal scaling and classical multivariate analysis. In particular a two-stage procedure which first scales the variables optimally, and then fits a simultaneous equations model, is studied in detail and is shown to have some desirable properties.
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  • 26
    ISSN: 1573-5036
    Keywords: organic matter ; rice ; submergence ; transformation ; zinc fraction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Laboratory and greenhouse experiments were conducted with two soilsviz., laterite and alluvial to study the transformation of applied Zn in soil fractions under submerged condition in the presence and absence of added organic matter and its relationship with Zn nutrition of rice plants. The results showed that application of organic matter caused a decrease in the concentration of Zn in shoot and root of rice plants and helped in translocating the element from root to shoot. The per cent utilization of applied Zn by plants was also found to increase by the application of organic matter. The transformation of applied Zn in different fractions in soils showed that a major portion (53.6–72.6%) of it found its way to mineral fractions leaving only 1.0–3.3, 6.6–18.9, 11.0–21.6 and 2.3–8.8% of the applied amounts in water soluble plus exchangeable, organic complexed, amorphous sesquioxides and crystalline sesquioxides bound fractions respectively. Application of organic matter favoured such transformation of applied Zn into these fractions except the mineral and crystalline sesquioxides bound ones. Simple correlation and multiple regression analyses between applied Zn in different soil fractions and fertilizer Zn content in plants showed that organic matter application increased the predictability of fertilizer Zn content in plants which has been attributed to the higher per cent recovery of applied Zn in plant available fractions in soils in presence of added organic matter.
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  • 27
    ISSN: 1573-5060
    Keywords: Beta vulgaris ; Beta procumbens ; Agrobacterium tumefaciens ; monosomic additions ; transformation ; chromosomal stability ; electrophoresis ; isozyme patterns ; in-vitro culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Crown galls were obtained from a monosomic addition plant of beet, carrying an extra chromosome of Beta procumbens, after transformation with a wild-type Agrobacterium tumefaciens strain. Analysis of peroxidase and esterase patterns obtained after cellulose acetate gel electrophoresis and iso-electric focusing respectively, gave evidence that no preferential loss of the alien chromosome on repeated subculture in-vitro from the crown galls occurred.
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  • 28
    ISSN: 1573-5060
    Keywords: Agrobacterium tumefaciens ; Beta vulgaris ; sugar beet ; regeneration ; shooter mutants ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Beta vulgaris plants were found to be susceptible to Agrobacterium tumefaciens strains carrying octopine Ti-plasmids after wounding of GA3 elongated stems or of hypocotyls. Tumors could be isolated and cultured aseptically. The tumor marker, octopine synthase (Ocs) activity, was present demonstrating the applicability of the Agrobacterium system for transfer of genetic information. For the production of transgenic plants two procedures were tested: inoculation of explants derived from cotyledons and hypocotyls of two weeks old seedlings and a leaf-disc procedure. The first method yielded both octopine positive calli as well as shoot regeneration on the six genotypes tested. In most cases, regeneration occurred from pre-existing, non-transformed meristems. The presence of Ocs activity could not be demonstrated in these shoots, although in one case octopine positive callus was formed at the base of the shoot, suggesting a chimeric structure of the plantlet or T-DNA genes, which were silent within the shoot and became active again in proliferating callus. The leaf-disc method did not give rise to direct or indirect regeneration, but transformed callus proliferated on the leaf edges. Optimal transformation frequencies were dependent on B. vulgaris genotype and Agrobacterium strain. The use of Agrobacterium shooter mutants or strains carrying an isolated cytokinin gene in order to influence endogenous phytohormone ratios did not result in the formation of shoots nor did it increase levels of regeneration in the first method. Further optimization is in order and in progress.
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  • 29
    ISSN: 1573-6822
    Keywords: hepatocytes ; hepatocarcinogenesis ; immunohistochemistry ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Monoclonal antibodies directed against antigens on rat liver epithelial cell lines were prepared. Three antibodies, 4C3, 19C6, and 3C2, recognized surface antigens present (although in different quantities) on eight epithelial cell lines tested, irrespective of whether they were normal or transformed. For MAb 3C2, the primary antigen common to all but one cell line showed a Mr of 135 kD. In paraffin sections of liver tissue, two antibodies, 40 and 19C6, reacted exclusively with bile duct epithelium, whereas the MAb 3C2 additionally reacted with sinusoidal endothelium and the endothelium of the portal venules. In sections of livers from rats exposed to diethylnitrosamine, the MAb 19C6 selectively stained bile duct-like structures in cholangiomas, while other preneoplastic and neoplastic lesions were not stained. These results demonstrate that the monoclonal antibodies obtained may prove useful for investigating cell lineages related to propagable liver epithelial cell lines and suggest that these cells may be derived from terminal bile ductular cells.
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  • 30
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    Journal of atmospheric chemistry 6 (1988), S. 47-59 
    ISSN: 1573-0662
    Keywords: Smog chamber ; terpenes ; alkanes ; alkenes ; reaction rate constants ; ozone ; OH radical ; photodegradation ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract The design and performance of a smog chamber for the study of photochemical reactions under simulated environmental conditions is described. The chamber is thermostated for aerosol experiments, and it comprises a gas chromatographic sample enrichment system suitable for monitoring hydrocarbons at the ppbv level. By irradiating NO x /alkane-mixtures rate constants for the reaction of OH radicals with n-alkanes are determined from n-pentane to n-hexadecane to be (k±2σ)/10−12 cm3 s−1=4.29±0.16, 6.2±0.6, 7.52 (reference value), 8.8±0.3, 10.2±0.3, 11.7±0.4, 13.7±0.3, 15.1±0.5, 17.5±0.6, 19.3±0.7, 22.3±1.0, and 25.0±1.3, respectively at 312 K. Rate constants, (k±2σ)/10−17 cm3 s−1, for the reaction of ozone with trans-2-butene (21.2±1.0), cis-3-methylpentene-(2) (47.2±1.7), cyclopentene (62.4±3.5), cyclohexene (7.8±0.5), cycloheptene (28.3±1.5), α-pinene (8.6±1.3), and β-pinene (1.4±0.2) are determined in the dark at 297 K using cis-2-butene (13.0) as reference standard.
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  • 31
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    Photosynthesis research 19 (1988), S. 23-37 
    ISSN: 1573-5079
    Keywords: binding ; uptake ; transformation ; DNA ; cyanobacteria ; etioplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Discoveries of the uptake and expression of various Escherichia coli plasmids by the cyanobacterium Anacystis nidulans and isolated cumber etioplasts are reviewed. In particular, the binding and uptake of nick-translated 32P-labeled plasmids and the expression of genes in the native plasmids are considered. Permeaplasts of A. nidulans 6301 and isolated EDTA-washed cucumber etioplasts exhibit binding and uptake of DNA that is unaffected by uncouplers of photophosphorylation or by dissipators of transmembrane proton graident. ATP inhibits both binding and udptake by permeaplasts or EDTA-washed etioplasts but the analog AMP-PNP (non-hydrolzable) is noninhibitory. With permeaplasts there is no effect of 20 mM Mg2+ (in the light) upon intake, whereas with EDTA-washed etioplasts, Mg2+ at the same concentration inhibits uptake as does 20 mM Ca2+. The transformation of A. nidulans 6301 to ampicillin-resistance by the plasmid pBR322 is much enhanced in permeaplasts. Indeed extracts of transformed cells catalyze the hydrolosis of the β-lactam nitrocefin. Transfromation of A. nidulans to antibiotic resistance may also be achieved with the plasmids pHUB4 and pCH1. The effect of light on transformation of A. nidulans 6301 differs with different plasmids. In pBR322 transformants the expression of ribulose bisphosphate carboxylase-oxygenase (RuBisCO) is markedly elevated. In these transformants, the foreign plasmid replicates by a pathway involving chromosomal integration and dissociation. The plasmid pCS75, a derivative of pUC9 (and therefore of pBR322) containing a Pst1 insert carrying genes for the large and small (S) subunits of RuBisCO from A. nidulans, is taken up and expressed in EDTA-washed cucumber cotyledon etioplasts. Expression is evidenced by the hydrolysis of nitrocefin and immunoprecipitation of labeled S subunits of RuBisCO (utilizing etioplasts which have been labeled with 35S-methionine after incubation with pCS75). The plasmid pUC9-CM carrying a cat gene is also expressed in cucumber etioplasts in a manner that demonstrates dependence both on the duration of etioplast washing by EDTA and plasmid concentration. Translation (as measured by 35S-methionine incorporation) by EDTA-washed etioplasts increases with cotyledon greening. However the enhancement of translation by prior incubation of EDTA-washed plastids with pCS75 decreases to zero during 24hr of cotyledon greening. Results suggest that the expression of foreign DNA in plastids may depend critically upon their developmental state.
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  • 32
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    Bulletin of experimental biology and medicine 105 (1988), S. 389-392 
    ISSN: 1573-8221
    Keywords: immortalization ; oncogenes ; transfection ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 33
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    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 37 (1988), S. 225-231 
    ISSN: 0730-2312
    Keywords: glycosyltransferase ; cell surface ; transformation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Recent studies have desmonstrated that Rous sarcoma virus-transformed baby hamster kidney (RS-BHK) cells express twofold higher levels of those N-linked oligosac-charides that contain the sequence [GlcNAc-β(1,6)Man (1,6)] compared to nontrans-formed parental BHK cells (Pierce and Arango, J. Biol. Chem. 261, 10772 [1986]). We have investigated in RS-BHK and BHK cells the activity of UDP-GlcNAc:α-D-mannoside β(l,6)N-acetylglucosaminyltransferase V, the enzyme that begins the synthesis of the sequence that is increased in the RS-BHK cells. We have measured GnT V activity using UDP- [3H]- GlcNAc and a synthetic oligosaccharide acceptor, GlcNAcβ(1,2)Man α(1,6)Manβ-O- (Ch2)8COOCH3, separating the radioactive product by a newly devised reverse-phase chromatographic technique. Assayed under optimal conditions, the specific activity of GnT V is about fourfold higher in RS-BHK sonicates than in BHK sonicates, suggesting that this increase in activity may be the primary mechanism that causes the increase in [GlcNAcβ(1,6) Man] sequences in the RS-BHK cells. The apparent Km, values of the enzymes in RS-BHK and BHK cell sonicates for UDP-GIcNAc and the synthetic acceptor are similar, as are the pH optima. These results suggest that the increase in GnT V-specific activity in RS-BHK cells is not caused by the presence in these cells of a GnT V with markedly different kinetic properties.
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  • 34
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    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 37 (1988), S. 61-78 
    ISSN: 0730-2312
    Keywords: heparan sulfate ; transformation ; cell surface proteoglycans ; growth control ; cancer glycosaminoglycans ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Cell surface proteoglycans are strategically positioned to regulate interactions between cells and their surrounding environment. Such interactions play key roles in several biological processes, such as cell recognition, adhesion, migration, and growth. These biological functions are in turn necessary for the maintenance of differentiated phenotype and for normal and neoplastic development. There is ample evidence that a special type of proteoglycan bearing heparan sulfate side chains is localized at the cell surface in a variety of epithelial and mesenchymal cells. This molecule exhibits selective patterns of reactivity with various constituents of the extracellular matrix and plasma membrane, and can act as growth modulator or as a receptor. Certainly, during cell division, membrane constituents undergo profound rearrangement, and proteoglycans may be intimately involved in such processes. The present work will focus on recent advances in our understanding of these complex macromolecules and will attempt to elucidate the biosynthesis, the structural diversity, the modes of cell surface association, and the turnover of heparan sulfate proteoglycans in various cell systems. It will then review the multiple proposed roles of this molecule, with particular emphasis on the binding properties and the interactions with various intracellular and extracellular elements. Finally, it will focus on the alterations associated with the neoplastic phenotype and will discuss the possible consequences that heparan sulfate may have on the growth of normal and transformed cells.
    Additional Material: 6 Ill.
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  • 35
    ISSN: 0730-2312
    Keywords: vanadate ; phosphotyrosine ; transformation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Rous sarcoma virus-trans formed baby hamster kidney fibroblasts (RSV/B4-BHK) adhere to a fibronectin-coated substratum by means of dot-like adhesion sites called podosomes in view of their shape and function as cellular feet (Tarone et al.: Exp Cell Res 159:141, 1985). Podosomes concentrate tyrosine-phosphorylated proteins, including pp60v-src, and appear in many cells transformed by oncogenes coding for tyrosine kinases. In this paper we used orthovanadate, an inhibitor of phosphotyrosine phosphatases, in order to increase the cellular concentration of phosphotyrosine and to study whether this treatment induced the cytoskeleton remodeling leading to the formation of podosomes. Indeed, orthovanadate (10-100 μM) induced in a time-and dose-dependent manner the redistribution of F-actin and the formation of podosomes in BHK cells. Cytoskeleton remodeling occurred along with a marked increase of tyrosine phosphorylatcd proteins. The vanadate effect on the cytoskeletal phenotype was enhanced by the simultaneous treatment of cells with a phorbol ester. Under the latter conditions almost all BHK cells showed podosomes. The vanadate effect was reversible insofar as podosomes and tyrosine-phosphorylated proteins disappeared. Then, vanadate treatment of normal cells induced the cascade of events leading to the cytoskeletal changes typical of transformation and suggested that the transformed cytoskeletal phenotype may he primarily induced by the tyrosine phosphorylation of unknown target(s) operated by endogenous kinases.
    Additional Material: 4 Ill.
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  • 36
    ISSN: 0192-253X
    Keywords: transformation ; gene structure ; cAMP ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cyclic nucleotide phosphodiesterase (phosphodiesterase) of Dictyostelium discoideum plays an essential role in development by hydrolyzing the cAMP used as a chemoattractant by aggregating cells. We have studied the biochemistry of the phosphodiesterase and a functionally related protein, the phosphodiesterase inhibitor protein, and have cloned the cognate genes. A 1.8-kb and a 2.2-kb mRNA are transcribed from the singlephosphodiesterase gene. The 2.2-kb mRNA comprises the majority of the phosphodiesterase mRNA found in differentiating cells and is transcribed only during development from a promoter at least 2.5 kb upstream of the translational start site. The 1.8-kb phosphodiesterase mRNA is detected at all stages of growth and development, is present at lower levels than the developmentally induced mRNA, and is transcribed from a site proximal to the protein-coding region. The phosphodiesterase gene contains a minimum of three exons, and a 2.3-kb intron, the longest yet reported for this organism. We have shown that the pds A. gene and fourfgd genes affect, the accumulation of the phosphodiesterase mRNAs, and we believe that these loci represent a significant portion of the genes regulating expression of the phosphodiesterase. The phosphodiesterase gene was introduced into cells by transformation and used as a tool to explore the effects of cAMP on the terminal stages of development. In cells expressing high levels of phosphodiesterase activity, final morphogenesis cannot be completed, and differentiated spore and stalk cells do not form. We interpret these results to support the hypothesis that cAMP plays an essential role in organizing cell movements in late development as well as in controlling the aggregation of cells in the initial phase of the developmental program.
    Additional Material: 6 Ill.
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  • 37
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    New York, NY [u.a.] : Wiley-Blackwell
    Applied Organometallic Chemistry 2 (1988), S. 303-307 
    ISSN: 0268-2605
    Keywords: Arsenic ; arsenic compounds ; transformation ; determination ; biological significance ; Chemistry ; Organic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Arsenic has a reputation as a poison, because arsenic trioxide was used during medieval times as an agent for murder. Lingering memories of these events make any arsenic-containing material suspect. Toxicity is a property of a specific compound and varies with the composition and structure of compounds. Developments in analytical methodology made it possible not only to determine total arsenic in a variety of matrices but also arsenic compounds. Knowledge about the arsenic cycle in marine systems has expanded considerably during the past decade. The marine arsenic cycle appears to be more complex than the cycle in the terrestrial environment. More attention must be given to the minor arsenic-containing compounds detected in organisms and experiments should be undertaken that provide information about the biochemical pathways used for the transformation of arsenic compounds.
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  • 38
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 9 (1988), S. 495-504 
    ISSN: 0192-253X
    Keywords: transformation ; extrachromosomal DNAs ; eukaryotic plasmids ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Cellular slime molds are one of only three types of eukaryotes known to contain circular nuclear plasmids. Unlike the 2-μm circle in Saccharomyces, different strains of Dictyostelium can carry different, nonhomologous plasmids. Covalently closed, circular DNA plasmids have been identified in D. discoideum, D. mucoroides, D. giganteum, and D. purpureum. These plasmids range in size from 1.3-27 kb and in copy number from 50-300 molecules per cell. Plasmids have been identified in approximately one-fifth of all isolates examined. The organization of their DNA in nucleosomes establishes their presence in the nucleus. We have successfully cotransformed endogenous Dictyostelium plasmids into D. discoideum using the G418 resistance shuttle vector B10S. Transformants carrying D. discoideum plasmids are recovered at much higher frequency than those carrying plasmids from the other Dictyostelium species. We have constructed recombinant plasmids based on the D. discoideum plasmid Ddp2 and the G418 resistance gene. With these extrachromosomal vectors, transformed cells are recovered at frequencies of up to 10-4 per input cell, the vectors are stably maintained at high copy number in the absence of selection, and the vectors can be used to introduce foreign DNA sequences into D. discoideum cells.
    Additional Material: 3 Ill.
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  • 39
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; cotton ; Gossypium hirsutum L. ; regeneration ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cotton (Gossypium hirsutum L.) cotyledon tissues have been efficiently transformed and plants have been regenerated. Cotyledon pieces from 12-day-old aseptically germinated seedlings were inoculated with Agrobacterium tumefaciens strains containing avirulent Ti (tumor-inducing) plasmids with a chimeric gene encoding kanamycin resistance. After three days cocultivation, the cotyledon pieces were placed on a callus initiation medium containing kanamycin for selection. High frequencies of transformed kanamycin-resistant calli were produced, more than 80% of which were induced to form somatic embryos. Somatic embryos were germinated, and plants were regenerated and transferred to soil. Transformation was confirmed by opine production, kanamycin resistance, immunoassay, and DNA blot hybridization. This process for producing transgenic cotton plants facilitates transfer of genes of economic importance to cotton.
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  • 40
    ISSN: 1573-4943
    Keywords: conformational energy ; three-dimensional structure ; amino acid substitutions ; P21 proteins ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The structural effects of amino acid substitutions at positions 12 and 16 in the amino-terminal segment (Tyr 4-Ala 18) of the ras-oncogene-encoded P21 proteins have been investigated using conformational energy analysis. The P21 protein with Val at position 12 and Lys at position 16 is known to have high transforming ability, while the P21 protein with Val at position 12 and Asn at position 16 is known to have poor transforming ability, similar to that of the normal protein (with Gly at 12 and Lys at 16.) The current results demonstrate a significant conformational change at position 15 induced by the substitution of Asn for Lys at position 16, which could explain this alteration in transformation potential. These findings are consistent with previous results suggesting the existence of a normal and a malignancy-causing conformation for the P21 proteins and suggest that the critical transforming region may encompass residues 12–15.
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  • 41
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    Potato research 30 (1987), S. 371-380 
    ISSN: 1871-4528
    Keywords: dihaploids ; protoclones ; protoplasts ; Solanum tuberosum ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Zusammenfassung Die Verwendung dihaploider Kartoffelklone (2n=2x=24) in Züchtungsprogrammen, vor allem die Rückkehr zum tetraploiden Grad, wird sehr oft durch deren fehlende Fertilität verhindert. Dieses Problem dürfte am elegantesten durch somatische Fusion der Dihaploiden überwunden werden. Als Ergebnis einer solchen Prozedur würden sich, ausser der Paarung beider Ploidiestufen, die Addition qualitativer und quantitativer Merkmale ergeben. Grösstes Hindernis für die Anwendung der Protoplastenfusion ist das Fehlen eines brauchbaren Selektionssystems für die Separierung von homo- und heterokaryotischen Fusionsprodukten. In der hier beschriebenen Methode wurde der sehr kräftige Wuchs einiger Kalli für die Vorselektion einiger vermuteter Hybriden verwendet. Nach Anwendung der Polyethylenglykol-Fusions-methode (PEG, Tabelle 1) konnten von fünf selektierten dihaploiden Klonen (P1–P5) grosse Zahlen von Kalli und Pflanzen regeneriert werden, obwohl die PEG-Behandlung einen negativen Einfluss auf die Regenerationsrate hatte (Tabelle 2). Insgesamt wurden nach PEG-Behandlung 115 Kalli als vermutliche Hybriden, ihrer extremen Wuchsleistung entsprechend, selektiert. Tabelle 3 vergleicht die Ploidiestufen dieser selektierten Klone mit der von unbehandelten Elternklonen. Wegen der somaklonalen Variation wurden auch von nicht fusionierten Protoplasten viele tetraploide Klone gefunden. Ihre Zahl war allerdings signifikant kleiner, und unter den nicht PEG-behandelten Protoplasten waren immer einige diploide vorhanden. Die Tabellen 4 und 5 zeigen die Merkmale von 9 und 10 selektierten Klonen (Hy 1–10) in vitro und in vivo für Sprosslänge, Zahl der Nodien, Blattfläche, Blattform, Zahl der Wurzeln und allgemeiner Wachstumsleistung. In allen Fällen waren die gemessenen Parameter bei den selektierten Klonen signifikant grösser als bei den Kontrollen. Folglich kann das stärkere Wachstum der selektierten Klone nicht nur mit somaklonaler Variation erklärt werden. Es ist ein starkes Indiz für die Hybridnatur. Das Isoenzym-Muster der Esterasen unterstreicht diese Schlussfolgerung. Den Ergebnissen zufolge ist es möglich, somatische Hybriden anhand ihrer hybriden Vitalität vorzuselektieren. Dies sollte die Möglichkeit zur Entdeckung somatischer Hybriden in ausreichender Häufigkeit für praktische Züchtungsprogramme erhöhen.
    Abstract: Résumé L'utilisation de clônes dihaploïdes (2n=2x=24) dans les programmes d'hybridation, et particulièrement le retour au niveau tétraploïde, est entravée par leur manque de fertilité. Ce problème pourrait être maitrisé élégamment par la fusion somatique de dihaploïdes. D'un tel procédé résulterait de plus l'héritage de caractères qualitatifs et quantitatifs apportés par le doublement de ploïdie. Le principal obstacle pour utiliser la fusion de protoplastes est l'absence d'un système de sélection approprié pour la séparation des homo et hétérokaryotes produits par la fusion. Par la méthode décrite dans cet article la grande vigueur de croissance de quelques cals a été mise à profit pour la présélection des présumés hybrides. Après l'adaptation du procédé de fusion au polyéthylène-glycol (PEG, tableau 1) sur cinq clônes dihaploïdes sélectionnés (P1–P5) un grand nombre de cals et de plantes pourrait être régénéré, bien que le traitement PEG ait une influence négative sur le taux de régénération (tableau 2). Au total 115 cals étaient sélectionnés après le traitement comme de présumés hybrides, en raison de leur extrème vigueur. Le tableau 3 compare les niveaux de ploïdie de ces clônes sélectionnés avec ceux des parents non traités. La variation somatique de protoplastes non fusionnés est également trouvée dans de nombreaux clônes tétraploïdes, leur nombre est cependant significativement plus petit, et parmi les protoclônes régénérés de protoplastes non traités quelques diploïdes sont toujours présents. Les tableaux 4 et 5 montrent les caractéristiques de 9 et 10 clônes sélectionnés (Hy 1–10) in vitro et in vivo, respectivement pour la longueur de pouses, le nombre de noeuds, la surface et la forme des feuilles, le nombre de racines et la vigueur générale. Dans tous les cas, les paramètres mesurés ont des valeurs significativement plus élevées dans les clônes sélectionnés que dans les témoins. En conséquence, leur vigoureuse croissance ne peut être expliquée seulement par la variation somatique mais elle constitue une indication sur la vigueur hybride. L'analyse des estérases souligne cette conclusion (figure 1). Ces résultats montrent qu'il est possible de présélectionner des hybrides somatiques par leur vigueur, ce qui augmente les possibilités de les détecter en quantité suffisante dans les programmes d'hybridation.
    Notes: Summary Tetraploid potato plants were regenerated after polyethylene-glycol-induced protoplast fusion between dihaploids. Hybrid vigour of the regenerated calli was used for preselection of fusion products. Nearly all the selected vigorous clones possessed chromosome counts at the tetraploid level. Fusion products were compared to the parental material to auto-fused plants of and to three protoclones expressing different degrees of somaclonal variation. The selected clones, where grown in vitro in growth rooms and in pots in the glasshouse, showed increased vigour compared to their parents, to auto-fused and to 4x protoclones. Plants of clones from very vigorous calli, when assessed by height, the number of nodes per plant, leaf morphology and tuber production, showed hybrid vigour. The hypothesis that superior clones result from heterokaryons after protoplast fusion or that they arise from other in vitro events such as somaclonal variation is discussed.
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  • 42
    ISSN: 1573-4935
    Keywords: c-Ha-ras ; metallothionein ; oncogene ; ras ; transformation ; tumorigenicity ; zinc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A C3H/10T1/2 cell line containing an inducible metallothionein-ras hybrid oncogene was conditionally and reversibly transformed upon exposure to zinc ions. Interestingly, although the cell line was fully malignant when expressing only low levels ofras, complete morphological transformation required much higher levels.
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  • 43
    ISSN: 1573-5028
    Keywords: geminivirus ; Ti-plasmid ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tomato golden mosaic virus (TGMV), a member of the geminivirus group, has a genome consisting of two DNA molecules designated the A and B components. Both are required for infectivity in healthy plants, although the former has been shown to replicate independently in transgenic plants containing tandem direct repeats of the A genome component. In the studies presented here, petunia plants transgenic for either both components (A×B hybrids) or the A component alone were examined for the presence of virus particles and encapsidated, single stranded viral DNA. The results of DNase protection experiments and direct observation of extracts from transgenic plants by electron microscopy indicate that single stranded TGMV DNA is in both cases packaged into paired particles identical to those obtained from virus-infected plants. DNase-treated virions isolated from A×B hybrid petunia are infectious when inoculated onto healthy Nicotiana benthamiana. Likewise, virions obtained from transgenic A petunia are infectious for plants transgenic for the B component. Our observations of TGMV replication in transgenic plants indicate that TGMV A DNA encodes all viral functions necessary for the replication and encapsidation of viral DNA. The possible role of the B component in TGMV replication is discussed.
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  • 44
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    Plant and soil 104 (1987), S. 175-181 
    ISSN: 1573-5036
    Keywords: rice ; soils ; submergence ; transformation ; zinc fractions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Distribution of different forms of Zn in 16 acid alluvial rice growing soils of West Bengal (India) and their transformation on submergence were studied. The results showed that more than 84% of total Zn occurred in the relatively inactive clay lattice-bound form while a smaller fractionviz. 1.1, 1.6, 11.1 and 2.0 per cent of the total occurred as water-soluble plus exchangeable, organic complexed, amorphous sesquioxide-bound and crystalline sesquioxide bound forms, respectively. All these four Zn forms showed significant negative correlations with soil pH (r=−0.48**, −0.39*, −0.61** and −0.67**, respectively), while the latter two Zn forms showed significant positive correlations with Fe2O3 (0.68** and 0.88***) and Al2O3 (0.89*** and 0.75***) content of the soils. The different Zn forms were found to have positive and significant correlations amongst each other, suggesting the existence of a dynamic equilibrium of these forms in soil. Submergence caused an increase in the amorphous sesquioxide-bound form of Zn and a decrease in each of the other three forms. The magnitude of such decreases in water-soluble plus exchangeable and crystalline sesquioxide-bound forms was found to be correlated negatively with initial pH values of the soils and positively with the increase in the amorphous sesquioxide-bound form, indicating their adsorption on the surface of the freshly formed hydrated oxides of Fe, which view was supported by the existence of significant positive correlation between the increase in the amorphous sesquioxide-bound form of Zn and that in AlCl3-extractable iron. The existence of a positive correlation between the decrease in crystalline sesquioxide-bound Zn and that in Fe2O3 content in soil suggested that on waterlogging the soil Zn occluded in the cry talline sesquioxide was released as a result of reduction of Fe2O3.
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  • 45
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    Plant cell, tissue and organ culture 9 (1987), S. 189-196 
    ISSN: 1573-5044
    Keywords: cereals ; genetic variability ; plant tissue culture ; somaclonal variation ; Sorghum bicolor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sorghum bicolor (L.) Moench, plants were regenerated from 4 to 5 month old callus cultures originally derived from seedling explants. Somaclonal variation was examined in the SC2 and SC3 generations of eight different clones plus the non-tissue cultured parent line. Characters examined included the area of the third leaf, height, tiller number, total shoot weight, seed number, grain yield, days to flowering, and chlorophyll content in the SC2 generation. Some plants were phenotypically different from the parent line in the SC2 generation. All clones were similar to the parent in third leaf area and chlorophyll content. Three clones had significant height reduction and six had significantly higher total plant matter production. Seven of the clones had significant increases in number of secondary tillers, and all had significant decreases in days to flowering. Only three clones had significant increases in grain yield and seed number; however, all the clones had smaller seeds. In subsequent studies of SC2 and SC3 generations of three somaclones, characters examined included height, tiller number, total shoot weight, and days to flower. In one of the somaclones none of the original differences in characters was maintained in two subsequent growing seasons in the SC2 or SC3 generation. The other two somaclones maintained the increased tiller number in the next growing season in both the SC2 and SC3 generations. In one of the somaclones plant height reduction occurred in the SC2 and SC3 generations. All other traits were not maintained.
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  • 46
    ISSN: 1573-5028
    Keywords: Agrobacterium rhizogenes ; binary vector ; kanamycin resistance ; transformation ; secondary metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Co-transfer of Agrobacterium rhizogenes T-DNA and T-DNA from the A. tumefaciens binary vector pBin19 (Bevan, 1984) was studied in detail using Nicotiana rustica. High frequencies of co-transfer of T-DNA's were observed, even when no selection pressure was exerted. Increased levels of pBin19 T-DNA were found in hairy root cultures with selection at higher levels of kanamycin sulphate (50–200 μg ml−1). Several other species were also transformed by A. rhizogenes carrying pBin19 and A. rhizogenes harbouring a different binary factor, pAGS125 (Van den Elzen et al., 1985), was used to transform N. rustica hairy roots to confer hygromycin B resistance.
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  • 47
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    Plant and soil 104 (1987), S. 143-146 
    ISSN: 1573-5036
    Keywords: decomposition ; ferulic acid ; lignin-related compounds ; mine soil microorganisms ; ring-fission products ; transformation ; vanillic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Spectrophotometric, chromatographic, and ion-exchange procedures were used to follow microbial transformation and degradation of lignin-related compounds during incubation in liquid culture. Analyses of solvent extracts from fermentation media showed that selected mine soil microorganisms were able to modify one or more of the structural components of vanillic and ferulic acids, including methoxyl groups, carbon side-chains, and aromatic rings.
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  • 48
    ISSN: 1573-5060
    Keywords: Solanum tuberosum ; potato ; amino acids ; amino acid analogue resistance ; frost tolerance ; plant regeneration ; proline ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A number of previously selected hydroxyproline (hyp) resistant cell lines of a diploid potato (Solanum tuberosum L., clone H2578, 2n=2x=24) could be regenerated into plants which were further analysed. Hyp resistance, although lower than in the originally selected calli, was still present in regenerated shoots and in callus initiated from these shoots and it was not lost upon (mini)tuber propagation. Regenerated shoots showed a wide range of phenotypic variation. The chromosome number, analysed in 4 clones, appeared to be hypotetraploid (44 or 45). Tuber-propagated regenerants generally showed increased frost tolerance both at the plant and the cell level. In leaves this seemed to be associated with increased levels of both proline and total amino acid content. However, in callus only the total amino acid content but not proline was still elevated. In the wild type the frost tolerance of the tubers appeared to be lower than that of the leaves. Between tubers of wild type and the hyp resistant regenerants no differences in frost tolerance were found, although proline and total amino acid content tended to be higher in tubers from the regenerants.
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  • 49
    ISSN: 1573-5028
    Keywords: acetosyringone ; Agrobacterium tumefaciens ; Arabidopsis thaliana ; leaf explants ; regeneration ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract High frequency transformation of Arabidopsis thaliana leaf explants has been obtained using a disarmed Ti plasmid containing the coding region of a neomycin phosphotransferase gene (NPT II) as a selectable marker. The rate of transformation ranged from 55 to 63 percent when acetosyringone (AS), a natural wound response molecule, was added to an Agrobacterium tumefaciens culture prior to incubation with leaf segments. Without acetosyringone, the transformation rate was approximately 2 to 3 percent. Calli resistant to G418 were regenerated into mature flowering plants in the presence of 10 μg/ml G418. Southern analysis and neomycin phosphotransferase assays confirmed the insertion and expression of the NPT II gene in regenerated Arabidopsis plants.
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  • 50
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; crown gall ; somaclonal variation ; T-DNA rearrangements ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract After three years of apparent stability in tissue culture, the single cell derived shooty crown gall line sNT1.013 produced a revertant shoot which had switched from non-rooting (Rod+) and octopine synthesizing (Ocs+) to Rod- Ocs-, indicating that in this revertant TL-DNA genes 4 (causing the Rod+ trait) and gene 3 (causing the Ocs+ trait) had been inactivated. Southern blots revealed that the inactivation of these T-DNA genes was the result of a considerable rearrangement of DNA sequences, accompanied by deletions and possibly also by DNA amplifications. This study for the first time unambiguously proves that foreign genes which have been introduced via Agrobacterium tumefaciens can, at a low frequency, be inactivated after T-DNA integration because of reorganization of T-DNA sequences during tissue culture. This can be considered as an event of somaclonal variation.
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  • 51
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    Breast cancer research and treatment 10 (1987), S. 217-227 
    ISSN: 1573-7217
    Keywords: breast cancer ; chromosome translocation ; gene amplification ; growth factor receptors ; neuroblastoma ; oncogenes ; prognosis ; proto-oncogenes ; small cell lung cancer ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
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  • 52
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    Plant molecular biology 8 (1987), S. 363-373 
    ISSN: 1573-5028
    Keywords: transformation ; MgCl2-PEG-electroporation ; competence ; protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Direct gene transfer into plant protoplasts has been recently developed, and conditions for high frequency transformation of SR1 tobacco protoplasts established. In this paper we analyse numerous transformation parameters in a comparative study on SR1 Nicotiana tabacum and N. plumbaginifolia, and report on a simple chemical technique for very efficient protoplast transformation. It is based on the synergistic interaction of MgCl2 and PEG. The technique yielded up to 1400 transformants per 3×105 treated N. tabacum protoplasts (up to 4.8% of the survivors, late selected clones). Using N. plumbaginifolia, the frequencies were 10-fold lower, indicating that the ‘competence’ for transformation has a species-specific component.
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  • 53
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    Euphytica 36 (1987), S. 659-665 
    ISSN: 1573-5060
    Keywords: Hordeum vulgare ; barley ; regeneration ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Plants were regenerated from callus cultures initiated from immature embryos of barley, Hordeum vulgare L. Immature embryos from seven diverse genotypes were cultured on modified Murashige and Skoog (MS) medium supplemented with 1.5 mg 2,4-D and 6.5 mg IAA/l. Of the 249 embryos cultured, 30% initiated callus within 8 days. Subculture of callus for 80 to 100 days on half-MS medium supplemented with 0.5 mg/l 2,4-D and 1.0 mg/l zeatin resulted in organogenesis. Culture of organogenic calli for 30 days on half-MS medium without growth regulators produced plants which originated mostly via multiple shoot formation. Callusing response of the tested genotypes ranged from zero to 44%; however, only 23% of the calli were regenerative. Regenerated plants included variants for chlorophyll deficiency, plant height, stem thickness, spike shape, pollen fertility, seed set and ploidy.
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  • 54
    ISSN: 1573-1561
    Keywords: Folivores ; marsupials ; allelochemicals ; transformation ; detox-ification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Measurements were made of the quantity and composition of the steam-volatile essential oils in gastrointestinal tract contents of greater gliders fedEucalyptus radiata foliage and brushtail possums fedE. melliodora foliage. In both species, there was less oil in the stomach contents than in an equivalent mass of foliage. Only minor losses of leaf oils occurred during mastication by greater gliders, and absorption from the stomach appeared to be the major reason for the difference in the oil content of ingested leaves and of stomach contents. The apparent digestibility of oils over the whole gut was 96–97 %, although oils from the cecum and feces of both species contained compounds not present in the original leaf oils. Absorption of oils before they reach the hindgut should reduce the severity of antimicrobial effects but may involve a metabolic cost to the animal in detoxification and excretion.
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  • 55
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    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 35 (1987), S. 51-68 
    ISSN: 0730-2312
    Keywords: steroid receptors ; heat shock protein ; transformation ; phosphorylation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: This brief review explores some recent observations relating to the structure of untransformed glucocorticoid and progesterone receptors and the mechanism by which the receptors are transformed to the DNA-binding state. In their molybdate-stabilized, untransformed state, progesterone and glucocorticoid receptors exist as a heteromeric 8-9S complex containing one unit of steroid binding phosphoprotein and one or two units of the 90 kD heat shock protein hsp90. When the receptors are transformed, the steroid-binding protein dissociates from hsp90. In cytosol preparations, temperature-mediated dissociation proceeds much more rapidly in the presence of hormone. The dissociated receptor binds to DNA with high affinity, regardless of whether it is in the hormone-bound or the hormone-free state. These observations raise the possibility that the primary, and perhaps the only, role for the hormone is to promote dissociation of the receptor-hsp90 complex.Molybdate, vanadate, and tungstate inhibit receptor transformation to the DNA-binding form, an effect that appears to reflect the ability of these transition metal oxyanions to stabilize the complex between the steroid receptor and hsp90. By promoting the formation of disulfide bonds, hydrogen peroxide also stabilizes the glucocorticoid receptor-hsp90 complex and prevents receptor transformation. A small, heat-stable factor present in all cytosol preparations inhibits receptor transformation, and, when the factor is removed, glucocorticoid receptors are rapidly transformed. This ubiquitous factor has the physical properties of a metal anion, and it is proposed that molybdate and vanadate affect steroid receptor complexes by interacting with a metal anion-binding site that is normally occupied by this endogenous receptor-stabilizing factor.
    Additional Material: 1 Ill.
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  • 56
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    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 33 (1987), S. 87-94 
    ISSN: 0730-2312
    Keywords: c-AMP ; G-proteins ; Ki-ras oncogene ; proliferation ; transformation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Rat kidney (NRK) cells infected with a temperature-sensitive mutant of the Kirsten sarcoma virus were arrested in the G0/G1phase of their cell cycle by incubation in serum-deficient medium at a p21-inactivating temperature of 41°C. These quiescent ts K-NRK cells were then stimulated to transit g1 and initiate DNA replication by lowering the temperature to 36°C, which rapidly reactivated p21 Reactivating the viral Ki-RAS protein by temperature shift led to an increase in adenylate cyclase activity in early G1 phase. The Ki-RAS protein increased the sensitivity of adenylate cyclase to guany1 nucleotides by a mechanism that seemed to involve inactivation of the enzyme's inhibitory G1regulatory protein.
    Additional Material: 4 Ill.
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  • 57
    ISSN: 0730-2312
    Keywords: myc-related genes ; nucleotide sequence ; transformation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The myc family of cellular oncogenes contains three well-defined members: c-myc, N-myc and L-myc. Additional structural and functional evidence now suggests that other myc-family oncogenes exist. The overall structure and organization of the c-, N-. and L-myc genes and transcripts are very similar. Each gene contains three exons: encoding a long 5′ untranslated leader and a long 3′ untranslated region. The proteins encoded by these myc genes share several stretches of significant homology. The conservation of sequences at the carboxy-terminus of the L-myc protein suggests that it is also a DNA-binding, nuclear-associated protein. Each myc gene will cooperate with an activated Ha-ras oncogene to cause transformation of primary rat embryo fibroblasts. Characteristics of several new myc-family members are described.
    Additional Material: 3 Ill.
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  • 58
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    Cellular and molecular life sciences 42 (1986), S. 1036-1038 
    ISSN: 1420-9071
    Keywords: Abelson virus ; centromere ; chromosome condensation ; premature chromosome condensation ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By fusing interphase cells to cells undergoing mitosis, the interphase chromosomes can be visualized. When analyzed in this way, chromosomes of normal mouse cells show characteristic undercondensed centromeric regions. We have found that the centromeric regions of chromosomes from Abelson virus-transformed cells are fully condensed. Abelson virus transforms mouse cells by introducing into them a virally encoded phosphokinase that is expressed constitutively. Thus, we propose that the condensation of centromeric chromatin is a result of overphosphorylation by the Abelson virus phosphokinase, and that the centromeric region is the relevant target of overphosphorylation in transformed cell growth.
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  • 59
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    Plant cell reports 5 (1986), S. 104-107 
    ISSN: 1432-203X
    Keywords: Plant tissue culture ; somaclonal variation ; mutable allele ; transposable element ; Medicago sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A white-flowered mutant (“WFM”) was regenerated from tissue culture of a purple-flowered plant of tetraploid alfalfa (Medicago sativa L.). When WFM was recultured, many regenerated plants (〉20%) were purple-flowered. Genetic analysis established that a functional allele, C2, of a locus required for anthocyanin pigmentation was in the simplex condition (C2c2c2c2) in the donor genotype when it mutated to an unstable recessive (“mutable”) allele, c2-m4, which is carried by WFM. Tissue culture experiments demonstrated that c2-m4 reverts to function at a high frequency in vitro. Results indicate that reversion occurs early in culture and may be the result of a genome shock associated with callus formation. Reversion also occurs in planta, but at a much lower frequency than in vitro. The c2-m4 allele is transmitted to progeny which revert in tissue culture. Revertant alleles, like the progenitor allele, are stable and are sexually transmitted. The action of a transposable element which is especially active in vitro is suggested.
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  • 60
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    Plant cell reports 5 (1986), S. 108-110 
    ISSN: 1432-203X
    Keywords: Plant tissue culture ; somaclonal variation ; mutable allele ; transposable element ; Medicago sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An unstable recessive (“mutable”) allele, c2-m4, of a locus required for anthocyanin pigmentation in alfalfa (Medicago sativa L.) reverts to a stable functional state at high frequency in vitro. It was previously established that a white-flowered mutant (“WFM”) and a white-flowered progeny of WFM (“WHGW3”) each carry the unstable allele. More than 20% of plants regenerated from tissue cultures of WFM and WHGW3 are revertant. It is here established that most nonrevertant plants regenerated from cultures of WFM and WHGW3 are stabilized in the recessive condition. Reculture of nonrevertants of WFM and WHGW3 indicated that there are three classes of nonrevertants: (i) Nonrevertants which revert in vitro at a high frequency typical of WFM; (ii) Nonrevertants which revert upon reculture but at significantly lower frequencies than WFM; and (iii) Nonrevertants which do not revert upon reculture. These observations are discussed in terms of transposable element action in vitro.
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  • 61
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    Bulletin of experimental biology and medicine 102 (1986), S. 1004-1007 
    ISSN: 1573-8221
    Keywords: erythrocytes ; transformation ; ultrastructure ; sensitization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 62
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    Methods in cell science 10 (1986), S. 165-172 
    ISSN: 1573-0603
    Keywords: transformation ; fibroblast ; C3H/10T1/2 ; mouse ; carcinogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Basic cell culture conditions required for execution of cell transformation studies with C3H/10T1/2 mouse embryo fibroblasts are described. These protocols permit assessment of the ability of chemicals to convert nontumorigenic C3H/10T1/2 cells to a tumorigenic phenotype characterized by altered controls upon in vitro cell division. Recent assay modifications that enhance the sensitivity of this focus formation assay and permit the study of multistage transformation processes are noted.
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  • 63
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    Methods in cell science 10 (1986), S. 157-164 
    ISSN: 1573-0603
    Keywords: BALB/c-3T3 cells ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The BALB/c-3T3 cell transformation assay evaluates the morphologic transforming potential of test chemicals using cultured mammalian cells as targets. The assay is semiquantitative and employs a contact-inhibited clone of cells derived from the original BALB/c-3T3 mouse cell line. Transforming activity, or a focus, is recognized as a dense layer of morphologically altered cells superimposed on a monolayer of the normal cell phenotype. The induction of transforming activity in a standard assay has been reported to correlate well with the carcinogenicity of many test chemicals in rodent bioassay; however, the standard assay does not detect carcinogens of all chemical classes. This report describes an operational protocol for the standard BALB/c-3T3 cell transformation assay. In addition, it provides assay acceptance criteria and a discussion of a method to evaluate transforming activity data.
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  • 64
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    Methods in cell science 10 (1986), S. 173-177 
    ISSN: 1573-0603
    Keywords: SHE ; clonal assay ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Procedures for the preparation of cryopreserved Syrian hamster embryo (SHE) cell pools and for the performance of the SHE clonal transformation assay are described. Attention to experimental details and the screening of cell pools and serum lots are important factors in obtaining consistent results with this clonal assay.
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  • 65
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    Methods in cell science 10 (1986), S. 185-188 
    ISSN: 1573-0603
    Keywords: transformation ; survival in aggregation ; carcinogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A sensitive and rapid transformation assay procedure is described. The end point, cell survival in aggregation, is measured by counting viable cells dissociated from aggregates in suspension for 4 d. When cells, previously treated with increased concentrations of a carcinogen, are suspended in liquid medium above an agar base, aggregates are formed, while untreated and solvent control cells remain primarily as single cells and show a rapid decline in cell survival; cells that had been treated with chemical carcinogen survive longer in suspension. When used as a marker for in vitro transformation, this end point can assess the carcinogenic potential of an agent.
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  • 66
    ISSN: 1573-0603
    Keywords: transformation ; human fibroblasts ; anchorage independence ; soft agar
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have determined the assay conditions necessary for quantifying the frequency of carcinogen-induced transformation of diploid human fibroblasts to anchorage independence. Emphasis is placed on the importance of using medium that supports clonal growth of the particular type of cells under investigation and titrating the percent of fetal bovine serum used in the assay to obtain a low, but measurable number of anchorage-independent colonies in the untreated control cultures. With this method, the target cells can be derived from adults as well as newborns and need not be in early passage, as long as they are able to grow vigorously.
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  • 67
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    Methods in cell science 10 (1986), S. 117-123 
    ISSN: 1573-0603
    Keywords: growth factors ; DNA synthesis ; cancer ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A rapid mitogenic assay suitable for the detection of transforming growth factors in the extracts of tissues or cells or in the medium conditioned by tumor cells in vitro is described. The method utilizes a nontumorigenic mouse embryo cell line (AKR-2B cells) maintained in serum-free conditions. Three classes of growth factors can be distinguished using this assay.
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  • 68
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    Plant cell, tissue and organ culture 6 (1986), S. 167-172 
    ISSN: 1573-5044
    Keywords: cape primrose ; Gesneriaceae ; anther culture ; somaclonal variation ; caulogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Regeneration of plants by caulogenesis of anther derived callus of Streptocarpus X hybridus ‘Concorde’ was obtained on MS basal medium with 2.0 mg/l NAA and 0.5 mg/l BAP. All regenerated plants were diploid and of somatic cell origin as determined by chromosome counts, flower color and morphological similarity to the anther donor plant, and flower color segregation among progeny of anther-derived plants. Variation expressed as deformed and dwarfed transient morphology was observed in 2 anther-derived plants. Variation was also observed among progenies of 5 anther-derived plants from the same anther donor plant for days to flowering, flowers per peduncle, peduncle length, and leaf area. Additional morphological variation including transient leaf variegation among seedlings and twisted or inverted leaves on mature plants was also observed among progeny of anther-derived plants.
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  • 69
    ISSN: 1573-5028
    Keywords: Agrobacterium tumefaciens ; A. rhizogenes ; hairy root ; plant ; transformation ; vector
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Binary Ti plasmid vector systems consist of two plasmids in Agrobacterium, where one plasmid contains the DNA that can be transferred to plant cells and the other contains the virulence (vir) genes which are necessary for the DNA transfer but are not themselves stably transferred. We have constructed two nononcogenic vectors (pARC4 and pARC8) based on the binary Ti plasmid system of Agrobacterium tumefaciens for plant transformation. Each vector contains the left and right termini sequences from pTiT37. These sequences, which determine the extent of DNA transferred to plant cells, flank unique restriction enzyme sites and a marker gene that functions in the plant (nopaline synthase in pARC4 or neomycin phosphotransferase in pARC8). After construction in vitro, the vectors can be conjugatively transferred from E. coli to any of several Agrobacterium strains containing vir genes. Using A. rhizogenes strain A4 containing the resident Ri plasmid plus a vector with the nopaline synthase marker, we found that up to 50% of the hairy roots resulting from the infection of alfalfa or tomato synthesized nopaline. Thus, vector DNA encoding an unselected marker was frequently co-transferred with Ri plasmid DNA to an alfalfa or a tomato cell. In contrast, the frequency of co-transfer to soybean cells was difficult to estimate because we encountered a high background of non-transformed roots using this species. Up to five copies of the vector DNA between the termini sequences were faithfully transferred and maintained in most cases suggesting that the termini sequences and the vir genes from the Ri and Ti plasmids are functionally equivalent.
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  • 70
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    Plant molecular biology 7 (1986), S. 43-50 
    ISSN: 1573-5028
    Keywords: transformation ; Agrobacterium tumefaciens ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Virulent strains of the soil bacterium Agrobacterium tumefaciens infect dicotyledonous plants and elicit a profound neoplastic response which results in crown gall formation (18). The inciting agent has been shown to be a high molecular weight plasmid (Ti) a section of which, the T-DNA, integrates into the host plant's genome (4, 28, 30). Although transformation of this kind was presumed to be limited to dicots, the detection of enzyme activities linked to the expression of T-DNA has been demonstrated in monocots from the families Liliaceae and Amaryllidaceae (10, 11). In this communication, we present evidence that a member of the commercially important Gramineae also is subject to A. tumefaciens directed transformation. This conclusion is based on two observations. First, seedlings of Zea mays that have had the bacteria introduced into wound sites defined by a region which includes the scutellar node and mesocotyl express the activity of enzymes whose synthesis is associated with the translation of T-DNA transcripts. Specifically, strain specific lysopine dehydrogenase activity has been detected in B6 infected material, whereas nopaline dehydrogenase activity is reported only in those plants inoculated with C58N. Second, the detection of either of these activities in extracts made from infected maize plants requires that the assaulting bacterial strain be competent with respect to the transfer of T-DNA. The vir - strains, JK195 and 238MX, are not, and transformation does not seem to occur. In this connection, the corresponding opine synthase activities are not observed.
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  • 71
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    Cell biology and toxicology 2 (1986), S. 9-20 
    ISSN: 1573-6822
    Keywords: amino acid transport ; α-aminoisobutyrate ; glutamate ; glutamine ; glutamine synthetase ; hormones ; primary hepatocyte cultures ; rat liver epithelial cells ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The capacities of Na+-dependent transport of α-aminoisobutyrate, glutamine and glutamate in four established and three transformed rat liver epithelial cell lines were found to be considerably higher than those of isolated and cultured hepatocytes. At least for transport systems A and G− this seemed to be due to elevated values of V max , whereas the values for K m were quite comparable to those of hepatocytes. In contrast to hepatocytes, however, no significant hormonal stimulation of amino acid uptake could be detected in the cell lines. Each normal cell line expressed a distinct pattern of transport capacities with respect to the three systems measured and this was not altered by chemical transformation of the lines. The individual patterns of the lines showed no similarity to presumptive patterns of subpopulations of liver parenchymal cells. In particular, there was no evidence for a direct relationship of one of the cell lines with a small subpopulation of parenchymal cells located adjacent to hepatic venules as revealed by additional measurements of glutamine synthetase, a marker enzyme for this particular subpopulation. It is concluded that established rat liver epithelial cell lines express features characteristic of normal hepatocytes with respect to amino acid transport, but have developed a distinct phenotype adapted to a rapid, hormone-independent growth in vitro. Alteration of their phenotype by transformation is not coupled with a further increase in amino acid transport capacity.
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  • 72
    ISSN: 1573-4943
    Keywords: conformational energy ; three-dimensional structure ; amino acid substitution ; P21 proteins ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The effects of amino acid substitutions for Gly 13 on the structure of the transforming region (Leu 6-Gly 15) of the P21 proteins have been explored using conformational energy calculations. It has been found that the substitution of Asp for Gly at this position results in a protein capable of transforming cells into malignant ones. Proteins that contain Ser at position 13 (but no other substitutions), however, transform cells with a greatly reduced activity. The transforming peptide with Asp 13 adopts a conformation that is different from the one for the peptide from the normal protein (with Gly 12 and Gly 13) and that may result in expression of a higher energy malignancy-producing form. The Ser-containing peptide adopts as its lowest energy conformation one that is identical to that of the peptide from the normal protein, thus explaining its lack of transforming activity. From analysis of the interactions preventing the Asp 13-containing peptide from adopting the “normal” conformation, it is predicted that substitutions of amino acids with branched side chains atC β, such as Val, Ile, and Thr, should promote cell transformation. This prediction with Val has recently been confirmed in genetic experiments.
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  • 73
    ISSN: 1573-4943
    Keywords: conformational energy ; three-dimensional structure ; amino acid substitution ; P21 proteins ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The conformational effects of different amino acid substitutions for Gly at position 12 in theras-oncogene-encoded P21 proteins have been investigated using conformational energy calculations. Mutations that cause amino acid substitutions for Gly 12 result in a protein that produces malignant transformation of cells. It had previously been shown that substitution of Val, Lys, or Ser for Gly at position 12 results in a major conformational change, and that the preferred lowest energy structure for each of the substituted peptides is identical. It is now found that substitution for Gly 12 of other amino acids that have widely disparate helix-nucleating potentials and completely different side chains (Asp, Asn, Cys, Phe, Tle, Leu, and Ala) all produce this identical lowest energy conformation. This finding is consistent with the recent results of site-specific mutagenesis experiments showing that P21 proteins containing these amino acids at position 12 all promote malignant transformation of cells and suggests the existence of a “malignancy-causing” conformation for the P21 proteins.
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  • 74
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    Euphytica 34 (1985), S. 525-537 
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; bread wheat ; somaclonal variation ; chromosomal aberrations ; wheat breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Transmission of somaclonal variation was studied in callus derived (SC1) plants of wheat, Triticum aestivum L. Second (SC2), third (SC3) and fourth (SC4) generations were compared with those obtained from embryo culture (E1-E4). SC2 generation had significantly lower grain yield (−37.3%), reduced thousand kernel weight (−7.8%) and shorter height (−2.1%) than that of E2, and included 5.7% seed-sterile and 15.0% partially seed-sterile plants. SC3 generation showed reduced yield (−12.6%), lower thousand kernel weight (−4.9%) and shorter plant height (−1.9%) than E3 generation. SC4 progeny of a short height, partially sterile SC1 variant included aberrant plants: a haploid, few aneuploids, mixoploids, some showing multiploidy, spindle abnormalities, multivalents, bridges and fragments, and tillers having supernumerary spikes with branched rachis. Somaclonal variation resulted from genome instability in callus, and likely involved transposable elements.
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  • 75
    ISSN: 1573-5060
    Keywords: Lotus corniculatus ; birdsfoot trefoil ; tissue culture ; somaclonal variation ; alteration stabiliy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Plants of Lotus corniculatus regenerated from single genotype-derived calli showed for several agronomic and morphological traits a variation similar to that present in the original population. Comparisons of the results obtained under field conditions during the 2nd and 3rd year from plant regeneration show: a) the maintenance of significant variance within callus-derived population for several traits examined; b) the total disappearance of ‘minus variants’ indicating a time-related decrease of depressive in vitro effects.
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  • 76
    ISSN: 1573-5028
    Keywords: Ti plasmid ; Hygromycin ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We show here that plant cells are sensitive to the antibiotic hygromycin-B4. We also show that a chimaeric gene consisting of the nopaline synthase (nos) gene regulatory elements and the E. coli derived hygromycin phosphotransferase (hpt) gene, when transferred to plants' cells, confers resistance to hygromycin B. The chimaeric nos-hpt gene enables efficient selection of DNA transfer to plant cells when used in conjunction with Ti plasmid-derived binary vectors in cocultivation experiments.
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  • 77
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    Plant cell, tissue and organ culture 4 (1985), S. 159-169 
    ISSN: 1573-5044
    Keywords: Plant tissue culture ; callus ; regeneration ; somaclonal variation ; isozymes ; chromosome instability ; celery ; Apium graveolens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genetically marked tissues of celery (Apium graveolens) were employed to contrast genetic and chromosomal stability in serially bulk-transferred callus and regenerated plants. After six months in culture, 84% of the callus cells were karologically indistinguishable from normal, while the remainder exhibited chromosome loss and/or fusion. All of 50 clones derived from this tissue expressed the control phenotype with respect to heterozygous isozyme markers. Of 95 plants regenerated from the same tissue, 94 were phenotypically indistinguishable from the original explant donor, and cytogenetic analyses revealed the presence in 4.3% of an accessory chromosome, while the remainder were normal diploids. Analysis of the selfed progeny of these regenerated plants revealed the presence of a new recessive mutation causing abnormal leaf morphology at a frequency of 1.8%. Only one of 40 cells in 12-month-old callus tissue was karyologically indistinguishable from normal, the remainder consisting primarily of hypodiploids. The observation that all 50 clones were phenotypically heterozygous was statistically inconsistent with the hypothesis that hypodiploidy was associated with random complete chromosome loss. The culture had, at this point, lost the ability to regenerate. It is speculated that embryogenic cloning of celery may be suitable under certain circumstances for direct field establishment, but that levels of new genetic variation are sufficiently high to preclude its use for seed production.
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  • 78
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    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 28 (1985), S. 1-6 
    ISSN: 0730-2312
    Keywords: epidermal growth factor ; 12-O-tetradecanoylphorbol-13-acetate ; EGF ; TPA ; promotion ; initiation ; transformation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Unlike 12-O-tetradecanoylphorbol-l3-acctate, epidermal growth factor (EGF) could not promote the appearance of type III foci from initiated C3H10T1/2 cells. At appropriate concentrations, EGF induced the formation of type II colonies in the absence of any initiator. At higher concentrations, EGF suppressed the induction of both type II and type III colonies elicited by methylcholanthrene.
    Additional Material: 3 Ill.
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  • 79
    ISSN: 1573-5060
    Keywords: Agrobacterium ; transformation ; lily ; β-glucuronidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Lily cv. Harmony was inoculated with several Agrobacterium strains to study its susceptibility to Agrobacterium infection and transformation. Tumorous tissue formation on inoculated stem internodes of sterile-grown plantlets, as well as expression of a β-glucuronidase marker gene interrupted by an intron in cells of inoculated stem nodes, indicate that the monocotyledon Lilium is a host for Agrobacterium.
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  • 80
    ISSN: 1573-5060
    Keywords: Hordeum vulgare ; isolated microspores ; particle bombardment ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A highly regenerable, isolated microspore system for barley, Hordeum vulgare L. cv. Igri, has been developed which is amenable to transformation studies using particle bombardment. The system allows DNA to be delivered to microspores at the single cell stage and both transient and stable transformation events have been demonstrated. The potential advantages of using isolated microspores as the target tissue in routine transformation systems are discussed.
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  • 81
    ISSN: 1573-5060
    Keywords: Lycopersicon ; tomato ; tomato spotted wilt virus ; tospovirus ; transformation ; virus resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Tomato spotted wilt virus (TSWV) causes significant economic losses in the commercial culture of tomato (Lycopersicon esculentum Mill.). Culture practices and introgression of natural sources of resistance to TSWV have only been marginally effective in controlling the TSWV disease. Recently however, high levels of protection against TSWV have been obtained by transforming tobacco with a chimaeric gene cassette comprising the TSWV nucleoprotein gene. This report demonstrates the successful application of this newly-created TSWV resistance gene in cultivated tomato. Transformation of an inbred tomato line with the TSWV nucleoprotein gene cassette resulted in high levels of resistance to TSWV that were maintained in hybrids derived from the parental tomato line. Therefore, transformant lines carrying the synthetic TSWV resistance gene make suitable progenitors for TSWV resistance to be incorporated into the breeding programmes of tomato.
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  • 82
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    Euphytica 85 (1955), S. 295-302 
    ISSN: 1573-5060
    Keywords: tissue culture ; somaclonal variation ; plant breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Somaclonal variation is a tool that can be used by plant breeders. The review examines where this tool can be applied most effectively and the factors that limit or improve its chances of success. The main factors that influence the variation generated from tissue culture are (1) the degree of departure from organised growth, (2) the genotype, (3) growth regulators and (4) tissue source. Despite an increasing understanding of how these factors work it is still not possible to predict the outcome of a somaclonal breeding programme. New varieties have been produced by somaclonal variation, but in a large number of cases improved variants have not been selected because (1) the variation was all negative, (2) positive changes were also altered in negative ways, (3) the changes were not novel, or (4) the changes were not stable after selfing or crossing. Somaclonal variation is cheaper than other methods of genetic manipulation. At the present time, it is also more universally applicable and does not require ‘containment’ procedures. It has been most successful in crops with limited genetic systems and/or narrow genetic bases, where it can provide a rapid source of variability for crop improvement.
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  • 83
    ISSN: 1573-5060
    Keywords: doubled haploids ; micropropagation ; mutant cultivars ; mutation techniques ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Conventional mutation techniques have often been used to improve yield, quality, disease and pest resistance in crops, or to increase the attractiveness of flowers and ornamental plants. More than 1700 mutant varieties involving 154 plant species have been officially released. In some economically important crops, e.g. barley, durum wheat and cotton, mutant varieties occupy the majority of cultivated areas in many countries. Mutation techniques have become one of the major tools in the breeding of ornamentals such as alstroemeria, begonia, chrysanthemum, carnation, dahlia and streptocarpus. The use of in vitro techniques such as anther culture, shoot organogenesis, somatic embryogenesis and protoplast fusion can overcome some of the limitations in the application of mutation techniques in both seed and vegetatively propagated crops. In vitro culture in combination with induced mutations can speed up breeding programmes, from the generation of variability, through selection, to multiplication of the desired genotypes. The expression of induced mutations in the pure homozygote obtained through microspore, anther or ovary culture, can enhance the rapid recovery of the desired traits. In some vegetatively propagated species, mutations in combination with in vitro culture technique, may be the only method of improving an existing cultivar. Currently, many molecular studies rely on the induction and identification of mutants in ‘model species’ for construction and subsequent saturation of genetic maps, understanding of developmental genetics and elucidation of biochemical pathways. Once identified and isolated, the genes that encode agronomically-important features can be either introduced directly into crop plants or used as probes to search for similar genes in crop species. It seems most likely that the recent developments based on these technologies will soon provide improved methods for selection of desired mutants.
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  • 84
    ISSN: 1573-5060
    Keywords: alfalfa ; alpha-amylase ; field performance ; manganese-dependent lignin peroxidase ; Medicago sativa ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Transgenic alfalfa plants expressinBacillus licheniformis alpha-amylase and mangaese-dependent lignin peroxidase (Mn-P) from Phanerochaete chrysosporium were produced using the Agrobacterium tumefaciens transformation system. In each case, there was a range of expression of the introduced gene among independent transgenic plants. Plants producing alpha-amylase showed no alteration of phenotype. Production of Mn-P in alfalfa, howeven, in most cases adversely affected plant growth and development. Affected plants were stunted with yellowing foliage, but survived and produced seed. Results from field trials showed that Mn-P production in transgenic alfalfa reduced dry matter yield and plant height. The extent of these symptoms and yield reduction was, for the most part, related to the level of foreign protein production as estimated by Western analysis. Field data from transgenic plants expressing alpha-amylase showed that there was no effect of foreign protein production on plant performance. Expression of Mn-P was shown to segregate in sexual progeny derived from transgenic plants.
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  • 85
    ISSN: 1573-5060
    Keywords: transformation ; silicon carbide ; whiskers ; maize
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary We review here the most recently developed technique for maize transformation which involves the vortexing of silicon carbide whiskers with maize cells in the presence of plasmid DNA. Fertile transgenic plants have been regenerated following whisker-mediated transformation which is compared with the alternatives described to date, namely protoplast uptake, particle bombardment and electroporation of intact tissue.
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  • 86
    ISSN: 1573-5060
    Keywords: Agrobacterium tumefaciens ; electroporation ; particle gun ; polyethylene glycol ; regeneration ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Suitable sunflower tissues and cells were transformed either by direct gene transfer into protoplasts, particle bombardment, or Agrobacterium co-culture. While all techniques allowed efficient short-term or transient expression of the introduced gene(s) in the respective tissues, stable transformation was only observed after transformation with Agrobacterium. The latter technique was suitable for the production of transgenic callus from seedling cotyledons and occasional shoots with chimaeric expression of the transgene. Detailed analysis of the interaction of Agrobacterium with this explant showed that infection efficiency was critically dependent on the co-culture conditions, and that the preferentially-transformed cells were not the ones competent for regeneration.
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  • 87
    ISSN: 1573-5060
    Keywords: Bacillus thuringiensis ; maize ; microprojectile bombardment ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A synthetic Bt gene encoding a truncated version of the CryIA(b) protein derived from Bacillus thuringiensis was successfully introduced into elite maize using microprojectile bombardment of immature embryos. The method used to initiate and identify transformation events is described. We describe the detailed parameters used for the Biolistics device as well as the plasmids used for the transformations. The plasmids contained the synthetic Bt gene driven by either the 35S CaMV promoter or a combination of two tissue-specific promoters, leaf and pollen, derived from maize. Specific conditions for the culture of Type I callus from immature embryos, the phosphinothricin (PPT) selection protocol, and the regeneration of plants are discussed. T0 and T1 plants were initially identified using the pH-dependent chlorophenol red test and/or the histochemical β-glucuronidase (GUS) assay. PCR and Southern data confirm the presence of the 35S CaMV promoter and the synthetic Bt gene.
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  • 88
    ISSN: 1573-5060
    Keywords: aluminium toxicity ; soil acidity ; somaclonal variation ; sorghum ; Sorghum bicolor (L.) Moench ; tissue culture ; salt stress ; drought stress ; variants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Sorghum bicolor (L.) Moench is generally quite sensitive to salt and acid (high aluminium) soil stresses, but quite tolerant of drought stress. As with any stress phenomenon, intra-specific variability exists within the genus. In vitro cell selection and somaclonal variation offer an alternative to traditional breeding methodology for generating improved breeding lines for hybrid development. A field selection protocol was developed for the three soil stresses and inter-stress evaluations were conducted in an effort to find multiple, stress-tolerant genotypes. The acid soil-drought stress, super-tolerant selections were located by the R7 generation when exposed to a combined aluminium-drought stress field environment and when the regeneration population (number of regenerated lines from one callus source) was maintained at 15,000 plants or higher. A variant frequency of 0.1 to 0.2% for stress tolerance and acceptable agronomic traits among the surviving somaclones, provided an adequate number of phenotypes with desirable agronomic characteristics and a high level of soil stress tolerance. Subsequent research verified that the stress-tolerant regenerants had superior acid soil and drought stress tolerance to that of the donor parents, that their yield capabilities under stress were superior to their parents, and that their stress tolerance attributes were transferred in hybrid combinations. In vitro selection was not effective in increasing the number of field stress survivors. In fact, superior germplasms were developed from non-stressed callus or salt-stressed callus. In vitro selection reduced regeneration frequency and subsequent survival of plants under field stress. In vitro-stressed regenerants should be subjected only to non-stressed environments to maintain population numbers for field selection and thereafter should be subjected to stress environments during later (R5+) generations. The optimal strategy for the exploitation of somaclonal variation may be through short-term cell culture (〈 12 months) with no attempt at in vitro selection.
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  • 89
    ISSN: 1573-5060
    Keywords: Brassica napus ; disease tolerance ; oxalic acid ; oxalate oxidase ; Sclerotinia sclerotiorum ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Oxalic acid is thought to have a primary role in the pathogenicity of several plant pathogens, notably Sclerotinia selerotiorum. A gene coding for the enzyme oxalate oxidase was isolated from barley roots and introduced into oilseed rape as a means of degrading oxalic acid in vivo. This report describes the production of several transgenic plants of oilseed rape and the characterisation of these plants by Southern, Western and enzyme activity assays. Plants were shown to contain an active oxalate oxidase enzyme and were tolerant of exogenously supplied oxalic acid.
    Type of Medium: Electronic Resource
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  • 90
    ISSN: 1573-5060
    Keywords: callus culture ; organogenesis ; pea ; Pisum sativum ; somaclonal variation ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The possibility of producing agronomically-useful somaclones via organogenesis and somatic embryogenesis from callus cultures of pea (Pisum sativum L.) was studied. Organogenic calli were induced from immature leaflets on MSB medium with NAA and BAP. Embryogenic calli were derived either from immature zygotic embryos (using 2,4-D) or from shoot apices (using picloram) of aseptically-germinated seedlings. The seed progenies (T1 to T3-generation) of primary regenerants were grown in field conditions and their phenotypic variation was evaluated and compared with control, non-tissue culture-derived plant material. In addition, electrophoretic analyses of selected isoenzyme systems and total proteins have been done. The results do not show dramatic changes in qualitative and quantitative traits. The evaluation of at least two future generations (T4, T5) is planned.
    Type of Medium: Electronic Resource
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  • 91
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 85 (1955), S. 1-12 
    ISSN: 1573-5060
    Keywords: genetic engineering ; gene targets ; mapping ; markers ; transformation ; QTLs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Type of Medium: Electronic Resource
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  • 92
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 85 (1955), S. 323-327 
    ISSN: 1573-5060
    Keywords: Brassica napus ; fatty acids ; gas chromatography ; Lunaria annua ; protoplast regeneration ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A programme of research was designed to investigate methods for the modification of the fatty acid profiles of high performance lines of oilseed rape (Brassica napus L.) in an attempt to produce lines with enhanced levels of industrially useful fatty acids. The methodology employed to achieve these objectives was based on the exploitation of somaclonal or protoclonal variation, and targeted somatic hybridization using wild cruciferous germplasm as fusion partners. A range of somaclonal lines was produced from shoot regeneration protocols. These lines underwent replicated, randomised glasshouse trials for morphological assessment followed by gas chromatographic analysis to monitor any changes in fatty acid profile. It was found that a small number of lines exhibited potentially useful changes in oleic acid and polyunsaturated fatty acid content. Protoplast regeneration and electrofusion protocols for a range of winter oilseed rape lines were developed, and methods for the isolation and fusion of protoplasts of the wild crucifer Lunaria annua (chosen for its high nervonic acid content) established.
    Type of Medium: Electronic Resource
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  • 93
    ISSN: 1573-5060
    Keywords: Linum usitatissimum ; linseed ; mutation breeding ; somaclonal variation ; fatty acids ; genetic engineering
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In the early 1980s the phenomenon of somaclonal variation induced by cell culture was exploited to produce genetic variation in linseed. The linseed variety Andro, derived from the widely grown Canadian variety McGregor, was selected in saline culture and was released for production in Canada. ‘Andro’ possesses traits very different from its parent, such as increased seedling vigour and tolerance to heat stress. Additional stable somaclonal variation in characters such as yield, days to maturity, seed weight and oil content were subsequently induced in ‘McGregor’. However, despite extensive screening of the somaclonal variants, no significant variation in the fatty acid profile was found. Chemical mutagenesis using ethyl methanesulphonate was, however, succesful in modifying the fatty acid profile of McGregor. Initial screening of M2 seed by the thiobarbituric acid colourimetric procedure was followed by gas chromatography to select half-seeds with atypical fatty acid profiles. Two independent, partially dominant genes were identified that were responsible for reducing the linolenic acid (18 : 3) from 50% to 2% while increasing linoleic acid (18 : 2) to 70%. A single, partially dominant gene, inherited independently of the linolenic acid genes, increased palmitic acid (16 : 0) from 7% to 30% and palmitoleic acid (16 : 1) from trace amounts to 4%. Agrobacterium-mediated transformation of linseed has also been successful. Herbicide tolerance genes for glyphosate, sulfonylurea and phosphinothricin have been incorporated into Canadian varieties. Commercially useful levels of tolerance to sulfonylurea herbicides have been achieved with no adverse agronomic affect. It is expected that a transgenic variety containing this resistance will be registered for commercial production in Canada in 1994. Standard breeding techniques, the application of antisense technology and the overexpression of fatty acid synthesis genes are being used to further modify the fatty acid profile of linseed, as well as for the transfer of abiotic stress-related genes identified in bromegrass.
    Type of Medium: Electronic Resource
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  • 94
    ISSN: 1573-5060
    Keywords: Agrobacterium ; plant regeneration ; potato ; Solanum tuberosum ; tissue culture ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary To provide a truly genotype-independent transformation system, it is necessary to be able to transform a wide range of potato genotypes. The ability to regenerate shoots in vitro was determined for 34 potato varieties using tuber disc explants. Following a culture regime used extensively in previous studies with the variety Desiree, half of the varieties could be regenerated from tuber discs and half could not. From a sample of varieties that could be regenerated from tuber discs, all but one variety gave transgenic plants. Twelve varieties were evaluated for the capacity to regenerate shoots from leaf and internode explants excised from in vitro grown plants. All of the varieties tested regenerated adventitious shoots. Leaf and internode explants from 5 varieties were subsequently used for transformation, and transgenic plants were produced from two potato varieties that did not give transgenic plants from tuber disc explants. Some varieties could not be transformed by either method, and will require modification of the in vitro regeneration and transformation system to be successful.
    Type of Medium: Electronic Resource
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  • 95
    Electronic Resource
    Electronic Resource
    Springer
    Euphytica 85 (1955), S. 131-134 
    ISSN: 1573-5060
    Keywords: apple ; transformation ; Agrobacterium ; preculture ; azacytidine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Leaf explants of apple cvs Gala and Golden Delicious were infected with the Agrobacterium tumefaciens strain AGL0(pMOG410). The effects of a 2 d preculture of the explants before infection and the addition of 5-azacytidine to the selection medium were studied. The percentages of GUS-positive explants after 5 w did not significantly alter due to these treatments. One of the ‘Gala’ shoots, which was removed from a leaf explant cultured for 8 w on selection medium, proved to be GUS-positive and will be analyzed further. In general, however, it should be concluded that regeneration of transgenic shoots directly from leaf tissue was not very effective.
    Type of Medium: Electronic Resource
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