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  • 201
    ISSN: 1573-6857
    Schlagwort(e): Ac/Ds ; transformation ; transgenic plants ; transposon tagging
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have recently shown that a plasmid-borneDissociation (Ds) element can excise from extrachromosomal plasmid DNA and integrate into a plant genome in the presence of theActivator (Ac) transposase.Ds andAc-carrying plasmids were used to co-transformNicotiana plumbaginifolia protoplasts. Transgenic plants were regenerated and analyzed. Here we describe further characterization of the system and discuss its efficiency in terms of DNA transformation and transposon tagging.
    Materialart: Digitale Medien
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  • 202
    ISSN: 1572-9702
    Schlagwort(e): Maternal microinjection ; transformation ; genetic improvement
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The persistence of DNA injected into two species of adult female phytoseiids and its transmission to serial eggs deposited by them was assessed by the polymerase chain reaction (PCR). The effect of DNA concentration on persistence and transmission was examined in Metaseiulus occidentalis. M. occidentalis females were microinjected with plasmid DNA at three different concentrations (250, 500, 750 ng μL−1) and allowed to deposit one to five eggs before the females and their last eggs were analyzed. Plasmid DNA was found in 82% of the females assayed and in 70% of all the eggs analyzed (including the fifth eggs produced after microinjection). Transmission of DNA to multiple eggs was also examined in Amblyseius finlandicus. Females of this species are less traumatized by microinjection allowing analysis of transmission over a more extended number of eggs. Females were microinjected and allowed to deposit eggs until their death. DNA from every fifth egg was analyzed by the PCR. PCR products were amplified from 51% of the eggs and from all egg classes except the 30th egg. The persistence and presence of plasmid DNA in both eggs and females suggests that (1) maternal microinjection is a more efficient method for DNA delivery than traditional egg microinjection, (2) it may be possible to isolate transformants from fewer maternally-microinjected females than originally expected, and (3) maternal microinjection could be useful as a DNA delivery system in other phytoseiids.
    Materialart: Digitale Medien
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  • 203
    Digitale Medien
    Digitale Medien
    Springer
    Antonie van Leeuwenhoek 65 (1994), S. 217-225 
    ISSN: 1572-9699
    Schlagwort(e): Gibberella fujikuroi ; gibberellins ; mutants ; regulation by nitrogen ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Gibberella fujikuroi (Fusarium moniliforme) is a complex group of plant pathogens. Some strains produce gibberellic acid and other gibberellins that promote growth and regulate various stages in plant development. The paper describes the research effort directed to development of genetic tools for this species. Furthermore the main features of the gibberellin biosynthetic pathway as established in Gibberella are described.
    Materialart: Digitale Medien
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  • 204
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 37 (1994), S. 39-46 
    ISSN: 1573-5044
    Schlagwort(e): endosperm cell culture ; maize ; protoplast ; transformation ; zeins
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Protoplasts were isolated from Zea mays (L.) A69Y endosperm suspension cultures and transformed by polyethylene glycol mediated DNA uptake with chimaeric gene constructs containing β-glucuronidase (GUS) or neomycin phosphotransferase II (NPTII); GUS-expressing and Kanamycin-resistant cultures were recovered. The transformed cells showed integration of the introduced foreign genes into genomic DNA and maintained their ability to synthesize endosperm-specific reserve proteins (zeins). No deletion or rearrangement of zein genes were observed in transformed cultures. Stable transformation of cultured maize endosperm cells may therefore represent a new methodological approach for the study of the transcriptional regulation of endosperm-expressed genes.
    Materialart: Digitale Medien
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  • 205
    ISSN: 1573-5044
    Schlagwort(e): monocotyledons ; Oryza sativa L. ; plant regeneration ; rice ; somatic embryogenesis ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract To obtain a reproducible efficient procedure for regeneration of rice plants through somatic embryogenesis from callus four published methods of callus induction and regeneration were compared. Callus was initiated from mature embryos of the Japonica cultivar Taipei 309 of rice (Oryza sativa L.). The number, mass and morphology of the callus formed on the scutellum were dependent on the medium used. A limited humidity and an optimal aeration of the culture vessels enhanced the frequency of embryogenesis and plant regeneration. A method described by Poonsapaya et al. (1989) was found to be the most efficient and was slightly modified. As a result 98% of the T309 embryos formed callus, of which 63% regenerated into plants. Each callus yielded an average of 6 plants. Plant morphology, fertility and seed set of the regenerants were found to be normal.
    Materialart: Digitale Medien
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  • 206
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 37 (1994), S. 257-269 
    ISSN: 1573-5044
    Schlagwort(e): adventitious shoots ; Malus x domestica Borkh. ; tissue culture ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Several factors that affect the frequency of organogenesis in apple leaf explants were examined for the scion cultivars ‘Empire’, ‘Freedom’, ‘Golden Delicious’, ‘Liberty’, ‘McIntosh’, and ‘Mutsu’ and for the rootstocks Malling 7A and Malling 26. The main factors affecting morphogenesis were BA concentration, basal medium, leaf explant origin and maturity, explant orientation, and photosynthetic photon flux. Depending on the genotype, optimal regeneration was obtained using either 22.2 or 31.1 μM BA and the N6 basal medium, with the exception of ‘Golden Delicious’ which regenerated better on MS medium. After 6 weeks, the average number of shoots per segment varied from 5 to 16, and the percentage of regeneration between 70 and 100%, depending on the genotype tested and the maturity of the explant. Regeneration capacity increased dramatically from the tip towards the base of the leaf, and was higher from the middle to the proximal end. Cefotaxime and carbenicillin, two antibiotics commonly used during transformation studies to eliminate Agrobacterium tumefaciens from plant tissue, were tested to determine their effect on morphogenesis. Cefotaxime at a dose of 250 mg 1-1 enhanced regeneration and shoot development, whereas carbenicillin at a dose of 500 mg l-1 induced abundant callus formation and inhibited regeneration. Kanamycin, a widely used selection agent for plant transformation, strongly inhibited regeneration even at very low doses. Schemes for selection and recovery of transgenic apple plants when kanamycin is used as the selection agent are discussed.
    Materialart: Digitale Medien
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  • 207
    Digitale Medien
    Digitale Medien
    Springer
    Plant growth regulation 15 (1994), S. 55-67 
    ISSN: 1573-5087
    Schlagwort(e): apple (Malus × domestica Borkh.) ; selection ; tissue culture ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Several factors that affect in vitro establishment, proliferation, and rooting of thirteen Malus cultivars and rootstocks were studied. Apple shoot tips (1.5±0.5 cm in length) were established using ascorbic and citric acids as antioxidants. Four proliferation media containing 1.0 mg 1−1 BA and different concentrations of IBA and GA3 were tested. Proliferation rates varied depending on the genotype and medium used. The highest proliferation rate was obtained for a rootstock that produced 11.6±2.5 shoots (1.5±0.8 cm in length) per tube per month. Rooting was induced with IBA for all the genotypes tested. The optimal IBA concentration was cultivar dependent (between 0.1 and 1.0 mg 1−1 IBA), and lower concentrations were necessary to induce rooting in liquid rather than in solid medium. The effects on shoot-tip proliferation of cefotaxime, carbenicillin and kanamycin, three antibiotics commonly used for transformation studies, were also evaluated. Cefotaxime at 200 mg 1−1 stimulated shoot growth and development, but at 500 mg 1−1 caused abnormal shoot morphology. Carbenicillin at 500 mg 1−1, alone or in combination with cefotaxime at 200 mg 1−1, inhibited proliferation and caused excessive enlargement of the basal leaves, inducing callus formation and release of phenolic compounds in the medium. Kanamycin at 50 mg 1−1 was phytotoxic and caused shoot chlorosis and necrosis. Consideration of the toxicity of these antibiotics is critical when designing transformation schemes for selection and recovery of transgenic apple plants.
    Materialart: Digitale Medien
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  • 208
    Digitale Medien
    Digitale Medien
    Springer
    Bulletin of experimental biology and medicine 118 (1994), S. 879-882 
    ISSN: 1573-8221
    Schlagwort(e): fibroblasts ; xenografts ; thymus-free animals ; transformation ; immortalization
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Abstract The tumorigenicity of cell clones derived from fibroblast lines isolated from colon cancer xenografts is studied in thymus-free animals. During cloning of the cell line obtained from the 3rd passage of the xenograft about 20% of the clones proved to be nontumorigenic, whereas such cells were not found in the line obtained from the 89th passage. Cytogenetic analysis of nontumorigenic clones revealed monosomy for the 13th chromosome with no alterations in the other chromosome pairs. Hybridization for the presence of Alu sequences was negative.
    Materialart: Digitale Medien
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  • 209
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 10 (1994), S. 141-149 
    ISSN: 0749-503X
    Schlagwort(e): Saccharomyces cerevisiae ; selectable marker ; transformation ; Life and Medical Sciences ; Genetics
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: Selectable markers integrated by the ‘gamma’ deletion method (Sikorski and Hieter, 1989) can be efficiently replaced in vivo with other markers by transformation with homologous plasmids. Transformation frequencies in experiments designed to replace original selectable markers with an alternate marker were high and molecular analysis confirmed that all transformants that exhibited the expected phenotypes (loss of the original prototrophy and gain of the alternate prototrophy) resulted from homologous recombination between plasmid sequences at the target locus. This technique involves no plasmid construction and greatly facilitates the generation of yeast cells containing multiple gene disruptions.
    Zusätzliches Material: 2 Ill.
    Materialart: Digitale Medien
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  • 210
    ISSN: 0268-2605
    Schlagwort(e): Arsenic ; methylation ; transformation ; freshwater food chain ; green alga ; shrimp ; killifish ; Chemistry ; Organic Chemistry
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Chemie und Pharmazie
    Notizen: Tolerance, bioaccumulation, biotransformation and excretion of arsenic compounds by the fresh-water shrimp (Neocaridina denticulata) and the killifish (Oryzias latipes) (collected from the natural environment) were investigated. Tolerances (LC50) of the shrimp against disodium arsenate [abbreviated as As(V)], methylarsonic acid (MAA), dimethylarsinic acid (DMAA), and arsenobetaine (AB) were 1.5, 10, 40, and 150μg As ml-1, respectively.N. denticulata accumulated arsenic from an aqueous phase containing 1 μg As ml-1 of As(V), 10 μg As ml-1 of MAA, 30 μg As ml-1 of DMAA or 150 μg As ml-1 of AB, and biotransformed and excreted part of these species. Both methylation and demethylation of the arsenicals were observed in vivo. When living N. denticulata accumulating arsenic was transferred into an arsenic-free medium, a part of the accumulated arsenic was excreted. The concentration of methylated arsenicals relative to total arsenic was higher in the excrement than in the organism.Total arsenic accumulation in each species via food in the food chainGreen algae (Chlorella vulgaris)→ shrimp (N. denticulata)→ killifish (O. latipes)decreased by one order of magnitude or more, and the concentration of methylated arsenic relative to total arsenic accumulated increased successively with elevation in the trophic level. Only trace amounts of monomethylarsenic species were detected in the shrimp and fish tested. Dimethylarsenic species in alga and shrimp, and trimethylarsenic species in killifish, were the predominant methylated arsenic species, respectively.
    Zusätzliches Material: 4 Ill.
    Materialart: Digitale Medien
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  • 211
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell reports 12 (1993), S. 468-473 
    ISSN: 1432-203X
    Schlagwort(e): Dioscorea alata ; GUS ; transformation ; particle gun
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary A biolistic particle gun was used to deliver genetic material into intact yam cells. Cultured suspension cells of D. alata were bombarded with microprojectiles coated with pBI221.2 DNA and histochemical assays were carried out to show transient GUS expression in bombarded cells. Stably transformed D. alata cells were recovered from cultured cells after bombardment with microprojectiles coated with pRT99gus harbouring both the nptII and uidA genes. Bombarded cells were selected on a medium containing geneticin (G418). Two months after bombardment, calli resistant to G418 were assayed for GUS expression. There was a 100% correlation between resistance to G418 and GUS expression. From these calli, four cell lines were established and GUS activity in each line was determined fluorometrically. The use of a specific GUS inhibitor showed that the GUS activity was due to the introduced uidA gene rather than to any intrinsic GUS-like activity originating from the plant. Incorporation of the introduced DNA into the plant genomic DNA was confirmed by Southern analysis.
    Materialart: Digitale Medien
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  • 212
    ISSN: 1432-203X
    Schlagwort(e): Acetosyringone ; Agrobacterium tumefaciens ; tomato ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Explants of tomato (Lycopersicon esculentum Mill cv. Ailsa Craig) were co-cultivated with Agrobacterium tumefaciens C58C1Rifr::pGSFR1161 in the presence of 20 (μM acetosyringone). Transformed root clones were selected on kanamycin medium and the presence of the nptII gene in the plant DNA confirmed by the polymerase chain reaction. Root clones derived from acetosyringone treatment grew more vigorously in the presence of kanamycin and synthesized a greater amount of NPT-II enzyme. The conclusion is that acetosyringone treatment enhances the transformation process, possibly by stimulating multiple insertions of the T-DNA into the host genome.
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  • 213
    ISSN: 1432-203X
    Schlagwort(e): Zea mays L ; microspore-derived cultures ; haploid ; regeneration ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Transgenic haploid maize (Zea mays L.) plants were obtained from protoplasts isolated from microspore-derived cell suspension cultures. Protoplasts were electroporated in the presence of plasmid DNA containing the gus A and npt II genes encoding ß-glucuronidase (GUS) and neomycin phosphotransferase II (NPT II), respectively. Transformed calli were selected and continuously maintained on kanamycin containing medium. Stable transformation was confirmed by enzyme assays and DNA. analysis. Stably transformed tissue was transferred to regeneration medium and several plants were obtained. Most plants showed NPT II activity, and some also showed GUS activity. Chromosome examinations performed on representative plants showed that they were haploid. As expected, these plants were infertile.
    Materialart: Digitale Medien
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  • 214
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 21 (1993), S. 937-942 
    ISSN: 1573-5028
    Schlagwort(e): α-tubulin ; Arabidopsis ; β-glucuronidase ; gene expression ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract In Arabidopsis tissues, the pool of tubulin protein is provided by the expression of multiple α-tubulin and β-tubulin genes. Previous evidence suggested that the TUA2 α-tubulin gene was expressed in all organs of mature plants. We now report a more detailed analysis of TUA2 expression during plant development. Chimeric genes containing TUA2 5′-flanking DNA fused to the β-glucuronidase (GUS) coding region were used to create transgenic Arabidopsis plants. Second-generation progeny of regenerated plants were analyzed by histochemical assay to localize GUS expression. GUS activity was seen throughout plant development and in nearly all tissues. The blue product of GUS activity accumulated to the highest levels in tissues with actively dividing and elongating cells. GUS activity was not detected in a few plant tissues, suggesting that, though widely expressed, the TUA2 promoter is not constitutively active.
    Materialart: Digitale Medien
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  • 215
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 21 (1993), S. 429-435 
    ISSN: 1573-5028
    Schlagwort(e): azacytidine ; DNA methylation ; gene expression ; inactivation ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have studied the effect of the demethylating agent azacytidine (azaC) on expression of a β-glucuronidase (GUS) gene transferred to tobacco leaf disks by Agrobacterium-mediated transformation. In a system where no selection was performed, where shoot formation was partially repressed, and where Agrobacterium does not express the GUS gene, we were able to follow the early events of transient and stable expression. Two days after inoculation, 8% of the cells expressed GUS but this proportion rapidly decreased to near zero in the following week. Treatment of leaf disks with azaC just after transformation retarded this inactivation to some extent, while treatment of Agrobacterium prior to transformation increased the frequency of transient expression. Three weeks after inoculation the number of GUS-expressing cells increased 4- to 6-fold in the leaf disks treated with azaC and in the leaf disks transformed with azaC-treated bacteria, while the control remained low. These data suggest that DNA methylation is involved in transgene inactivation and that a large number of silent but potentially active transgenes become integrated.
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  • 216
    ISSN: 1573-5028
    Schlagwort(e): Zea mays L. ; protoplast ; DNA uptake ; transformation ; β-glucuronidase ; promoter ; α-amylase gene
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A reproducible and efficient transformation system has been developed for maize that is based on direct DNA uptake into embryogenic protoplasts and regeneration of fertile plants from protoplast-derived transgenic callus tissues. Plasmid DNA, containing the β-glucuronidase (GUS) gene, under the control of the doubled enhancer element (the −208 to −46 bp upstream fragment) from CaMV 35S promoter, linked to the truncated (up to −389 bp from ATG) promoter of wheat, α-amylase gene was introduced into protoplasts from suspension culture of HE/89 genotype. The constructed transformation vectors carried either the neomycin phosphotransferase (NPTII) or phosphinothricin acetyltransferase (PAT) gene as selective marker. The applied DNA uptake protocol has resulted at least in 10–20 resistant calli, or GUS-expressing colonies after treatment of 106 protoplasts. Vital GUS staining of microcalli has made possible the shoot regeneration from the GUS-stained tissues. 80–90% of kanamycin or PPT resistant calli showed GUS activity, and transgenic plants were regenerated from more than 140 clones. Both Southern hybridization and PCR analysis showed the presence of introduced foreign genes in the genomic DNA of the transformants. The chimeric promoter, composed of a tissue specific monocot promoter, and the viral enhancer element specified similar expression pattern in maize plants, as it was determined by the full CaMV 35S promoter in dicot and other monocot plants. The highest GUS specific activity was found in older leaves with progressively less activity in young leaves, stem and root. Histochemical localization of GUS revealed promoter function in leaf epidermis, mesophyll and vascular bundles, in the cortex and vascular cylinder of the root. In roots, the meristematic tip region and vascular tissues stained intensively. Selected transformants were grown up to maturity, and second-generation seedlings with segregation for GUS activity were obtained after outcrossing. The GUS-expressing segregants carried also the NPTII gene as shown by Southern hybridization.
    Materialart: Digitale Medien
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  • 217
    ISSN: 1573-5028
    Schlagwort(e): transformation ; promoters ; introns ; gene expression ; Oryza sativa ; Hordeum vulgare ; Lolium perenne
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Transcriptional and translational fusions were made between the reading frame coding for β-D-glucuronidase and sequences of either a constitutively expressed rice gene (GOS2) involved in initiation of translation or a light-inducible rice gene (GOS5). The transient expression of the fusions was studied via particle bombardment of seedling tissues of rice, perennial ryegrass and barley. Furthermore, the results of transient and stable expression were compared for cell suspensions of four rice varieties, one barley variety and one perennial ryegrass variety. TheGOS2-gusA fusions were active in all three monocots studied. Best results were obtained for a construct having both a transcriptional and a translational fusion as well as intron and exon sequences (PORCEHyg). The level of GUS activity was in the range of activities as obtained by the 35S CaMV promoter transcriptionally fused togusA. ThegusA fusion with the light-inducible gene (GOS5) was active in green seedling tissues of all monocots studied. Also a weak expression compared to theGOS2 constructs was found in stably transformed rice callus. ThegusA fusions with the mannopine synthase promoters 1′ and 2′ of the TR-DNA were transiently expressed at lower levels in cell suspensions than PORCEHyg. For stably transformed rice callus the expression of theGOS2-gusA fusion often decreased during prolonged subculture. This decrease in GUS activity and the various GUS-staining phenotypes of transgenic calli are explained by the presence of different cell types in the suspensions used and in the calli. It is presumed that the nature of the cells and their relative contribution in the calli change drastically upon further subculture.
    Materialart: Digitale Medien
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  • 218
    ISSN: 1573-5028
    Schlagwort(e): transformation ; promoters ; introns ; gene expression ; Oryza sativa ; Hordeum vulgare ; Lolium perenne
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Transcriptional and translational fusions were made between the reading frame coding for β-D-glucuronidase and sequences of either a constitutively expressed rice gene (GOS2) involved in initiation of translation or a light-inducible rice gene (GOS5). The transient expression of the fusions was studied via particle bombardment of seedling tissues of rice, perennial ryegrass and barley. Furthermore, the results of transient and stable expression were compared for cell suspensions of four rice varieties, one barley variety and one perennial ryegrass variety. The GOS2-gusA fusions were active in all three monocots studied. Best results were obtained for a construct having both a transcriptional and a translational fusion as well as intron and exon sequences (PORCEHyg). The level of GUS activity was in the range of activities as obtained by the 35S CaMV promoter transcriptionally fused to gusA. The gusA fusion with the light-inducible gene (GOS5) was active in green seedling tissues of all monocots studied. Also a weak expression compared to the GOS2 constructs was found in stably transformed rice callus. The gusA fusions with the mannopine synthase promoters 1′ and 2′ of the TR-DNA were transiently expressed at lower levels in cell suspensions than PORCEHyg. For stably transformed rice callus the expression of the GOS2-gusA fusion often decreased during prolonged subculture. This decrease in GUS activity and the various GUS-staining phenotypes of transgenic calli are explained by the presence of different cell types in the suspensions used and in the calli. It is presumed that the nature of the cells and their relative contribution in the calli change drastically upon further subculture.
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  • 219
    ISSN: 1573-5028
    Schlagwort(e): bar ; herbicide resistance ; phosphinothricin acetyltransferase ; rice ; selectable marker ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have used the bar gene in combination with the herbicide Basta to select transformed rice (Oryza sativa L. cv. Radon) protoplasts for the production of herbicide-resistant rice plants. Protoplasts, obtained from regenerable suspension cultures established from immature embryo callus, were transformed using PEG-mediated DNA uptake. Transformed calli could be selected 2–4 weeks after placing the protoplast-derived calli on medium containing the selective agent, phosphinothricin (PPT), the active component of Basta. Calli resistant to PPT were capable of regenerating plants. Phosphinothricin acetyltransferase (PAT) assays confirmed the expression of the bar gene in plants obtained from PPT-resistant calli. The only exceptions were two plants obtained from the same callus that had multiple copies of the bar gene integrated into their genomes. The transgenic status of the plants was varified by Southern blot analysis. In our system, where the transformation was done via the protoplast method, there were very few escapes. The efficiency of co-transformation with a reporter gene gusA, was 30%. The To plants of Radon were self-fertile. Both the bar and gusA genes were transmitted to progeny as confirmed by Southern analysis. Both genes were expressed in T1 and T2 progenies. Enzyme analyses on T1 progeny plants also showed a gene dose response reflecting their homozygous and heterozygous status. The leaves of To plants and that of the progeny having the bar gene were resistant to application of Basta. Thus, the bar gene has proven to be a useful selectable and screenable marker for the transformation of rice plants and for the production of herbicide-resistant plants.
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  • 220
    ISSN: 1573-5028
    Schlagwort(e): carboxysomes ; cyanobacteria ; rbc genes ; Rubisco ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The cyanobacterium Synechocystis PCC6803 was chosen as a target organism for construction of a suitable photosynthetic host to enable selection of variant plant-like ribulose bisphosphate carboxylase/oxygenase (Rubisco) enzymes. The DNA region containing the operon encoding Rubisco (rbc) was cloned, sequenced and used for the construction of a transformation vector bearing flanking sequences to the rbc genes. This vector was utilized for the construction of a cyanobacterial rbc null mutant in which the entire sequence comprising both rbc genes, was replaced by the Rhodospirillum rubrum rbcL gene linked to a chloramphenicol resistance gene. Chloramphenicol-resistant colonies, Syn6803†rbc, were detected within 8 days when grown under 5% CO2 in air. These transformants were unable to grow in air (0.03% CO2). Analysis of their genome and Rubisco protein confirmed the site of the mutation at the rbc locus, and indicated that the mutation had segregated throughout all of the chromosome copies, consequently producing only the bacterial type of the enzyme. In addition, no carboxysome structures could be detected in the new mutant. Successful restoration of the wild-type rbc locus, using vectors bearing the rbc operon flanked by additional sequences at both termini, could only be achieved upon incubating the transformed cells under 5% CO2 in air prior to their transferring to air. The yield of restored transformants was proportionally related to the length of those sequences flanking the rbc operon which participate in the homologous recombination. The Syn6803Δrbc mutant is amenable for the introduction of in vitro mutagenized rbc genes into the rbc locus, aiming at the genetic modification of the hexadecameric type Rubisco.
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  • 221
    ISSN: 1573-5028
    Schlagwort(e): cis elements ; light regulation ; Rca promoter ; rubisco activase ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Deletions in the spinach rubisco activase (Rca) promoter in transgenic tobacco were analyzed to define the regions necessary for conferring light-inducible and tissue-specific expression. Transgenic plants were constructed with Bal 31 deletions of the Rca promoter fused to the coding region of the bacterial reporter gene β-glucuronidase (GUS). Analysis of the Rca deletion mutants localized the region conferring normal expression downstream from −294 relative to the Rca transcription start site. A second set of transgenic plants containing the cauliflower mosaic virus (CaMV) 35S enhancer fused to the 3′ end of the Rca/GUS constructs demonstrated the presence of a light-responsive element between −150 and −78 active in leaves. Regions 10 bp long within the light-responsive region, which included putative G box and GT elements, were removed by recombinant polymerase chain reaction. Deletion of the G box element resulted in a loss of gene expression in the leaves of transgenic tobacco, while deletion of the GT motif caused a 10–100-fold increase in expression in roots. However, site-directed mutagenesis of the GT motif resulted in expression patterns identical to the normal promoter. These experiments demonstrated that light-inducible and tissue-specific expression of the Rca promoter involves multiple cis elements proximal to the transcription start site, and that interactions between these elements are essential for regulating expression.
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  • 222
    ISSN: 1573-5044
    Schlagwort(e): Agrobacterium ; Dendranthema grandiflora ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract In an attempt to develop a system for producing transformed plants from explants ofDendranthema grandiflora, the susceptibility of the cultivar Super White to various wild-type strains ofAgrobacterium tumefaciens andA. rhizogenes was investigated. Tumour formation was not a reliable indicator of the ability of a related disarmed strain to mediate transformation. Following inoculation of explants with disarmedAgrobacterium strains, a number of shoots developed on selective media. However, none of these shoots were transformed. By co-cultivating stem internode explants with a mixed inoculum of wild-type and disarmed strains, it was possible to obtain a callus stably transformed withAgrobacterium carrying a disarmed T-DNA. Histological analysis of explants revealed that shoot regeneration initially occurred from the cells of the epidermis and subsequently from the cortex. However, the cells which were susceptible to T-DNA transfer were confined to the vascular tissue.
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  • 223
    Digitale Medien
    Digitale Medien
    Springer
    Plant growth regulation 13 (1993), S. 77-84 
    ISSN: 1573-5087
    Schlagwort(e): Agrobacterium rhizogenes ; auxin ; indole-3-acetic acid (IAA) ; root initiation ; sensitivity ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract This paper is the second part of a review which considers evidence for the involvement of auxin in root initiation. Part II examines the research being carried out with transformed plant tissues. Agrobacterium rhizogenes causes abundant root initiation at the site of inoculation. Ri plasmid T-DNA contains several genes which encode enzymes involved in the biosynthesis and metabolism of indole-3-acetic acid. Transfer of various fragments of the Ri plasmid has also been reported to confer increased sensitivity to auxin upon plant cells. Controlled expression of these genes in the plant genome potentially offer an insight for developmental plant physiologists into the role of plant growth substances in the process of root initiation. The importance of absolute levels of IAA in the stimulation of root initiation is discussed.
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  • 224
    ISSN: 1573-6857
    Schlagwort(e): plant ; genetic engineering ; nutritive value ; agrobacterium ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract This review describes work aimed at the improvement of the nutritive value of grain and forage legumes using gene transfer techniques. Two traits which are amenable to manipulation by genetic engineering have been identified. These are plant protein quality and lignin content. In order to increase the quality of protein provided by the legume grains peas and lupins, we are attempting to introduce into these species chimeric genes encoding a sunflower seed protein rich in the sulphur-containing amino acids methionine and cysteine. These genes are designed to be expressed only in developing seeds of transgenic host plants. Chimeric genes incorporating a similar protein-coding region, but different transcriptional controls, are being introduced into the forage legumes lucerne and subterranean clover. In this case the genes are highly expressed in the leaves of transformed plants, and modifications have been made to the sunflower seed protein-coding sequences in order to increase the stability of the resultant protein in leaf tissue. Another approach to increasing plant nutritive value is represented by attempts to reduce the content of indigestible lignin in lucerne.
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  • 225
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 32 (1993), S. 263-270 
    ISSN: 1573-5044
    Schlagwort(e): Dendranthema grandiflora ; preculture ; regeneration ; transformation ; β-glucuronidase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Explants from leaves of in vitro-grown chrysanthemum (Dendranthema grandiflora Tzvel.) cultivars regenerated adventitious shoots without an intermediate callus phase. Puncturing explants with a brush increased regenerations, but in combination with cocultivation with Agrobacterium tumefaciens it had an adverse effect on shoot formation. The negative effect of brushing and cocultivation could be overcome by preculturing explants for 8 days. Preculture altered the location of transformed sites but did not inhibit transformation. Regeneration following cocultivation with Agrobacterium is also encouraged if alternative regeneration protocols are used that do not require brushing.
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  • 226
    Digitale Medien
    Digitale Medien
    Springer
    Euphytica 71 (1993), S. 1-14 
    ISSN: 1573-5060
    Schlagwort(e): Agrobacterium tumefaciens ; biolistics ; co-suppression ; co-transformation ; electroporation ; epistasis ; gene silencing ; somaclonal variation ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary The DNA delivery systems which are routinely used to introduce genes into crop plants are Agrobacterium tumefaciens, electroporation and particle bombardment. The differences and similarities between these different transformation techniques are outlined. The influence of the cell biological approach, and more specifically the impact of the state of the plant cell at the moment of transformation, on the genotype and phenotype of the regenerated transgenic plant is analysed. In this respect phenomena such as position effects, gene silencing, co-suppression, epistasis, co-transformation and somaclonal variation are discussed. The relevance of these factors for plant breeders is discussed.
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  • 227
    Digitale Medien
    Digitale Medien
    Springer
    Journal of inclusion phenomena and macrocyclic chemistry 15 (1993), S. 27-36 
    ISSN: 1573-1111
    Schlagwort(e): Crystallization ; nucleation ; crystal growth ; polymorph ; molecular complex ; transformation ; solvent effect
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Chemie und Pharmazie
    Notizen: Abstract The effect of the solvents acetone (AT), dimethylsulfoxide (DMSO) and methylcellosolve (MCS) on the inclusion of 2-acetylnaphthalene (2-AN) in the host, 1,1-di(p-hydroxyphenyl)cyclohexane (DHC) has been investigated. Each solvent molecule is included in DHC in a molar ratio of 1.0, when DHC is crystallized from the solvents. The evaporation rate of these solvents from the host lattice decreases in the order AT, MCS and DMSO. The order agrees well with the interaction strength between the host and solvent molecule, which was measured by DSC and IR. 2-AN cannot be included in the crystals by crystallization from MCS and DMSO solutions. However, in AT solution both AT and 2-AN are included competitively and the morphology of the crystals is different from that obtained in pure solution. The amount of 2-AN in the crystals increases continuously with its concentration in solution. This behavior indicates that AT is replaced by 2-AN and the solid solution of the molecular complex is formed. The solid solution is a metastable form and the solution-mediated tranformation to the stable form (which includes only AT) was observed.
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  • 228
    ISSN: 1573-9368
    Schlagwort(e): Peanut ; Arachis hypogaea ; transformation ; callus ; Agrobacterium tumefaciens ; peanut stripe virus coat protein
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Transformed callus was produced from peanut (Arachis hypogaea L. cv. Okrun) hypocotyl explants after four days of co-cultivation withAgrobacterium tumefaciens strains EHA101, LBA4404 or ASE1 carrying the binary vector pKYLX71GUS on a defined medium followed by selection with kanamycin (200 mg l−1). Transformed calluses were cultured as independent cell lines potentially derived from a single transformation event. Stable integration and expression of foreign gene(s) in the callus was confirmed by Southern and western blot analyses and enzyme assays. A few cell lines showed a single insert of the foreign gene. Using the above protocol, transformed peanut callus expressing the peanut stripe virus coat protein gene was obtained.
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  • 229
    ISSN: 1573-9368
    Schlagwort(e): Lotus ; Agrobacterium rhizogenes ; transformation ; hygromycin resistance ; tannins
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The speciesLotus corniculatus andL. tenuis were transformed with anAgrobacterium rhizogenes binary vector, conferring resistance to the antibiotic hygromycin. Transgenic plants recovered from both species were tested for the ability of leaf-derived calluses to grow in a hygromycin-supplemented medium. Molecular analysis showed the integration of the Ri T-DNA and of the gene for hygromycin resistance, with a high frequency of co-transformation. Progeny analysis of the hygromycin resistance indicated this to be a single Mendelian trait in test plants. The transformed plants will be utilized in somatic hybridization experiments with lucerne for producing non-bloating genotypes with condensed tannins in leaves.
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  • 230
    Digitale Medien
    Digitale Medien
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 53 (1993), S. 61-73 
    ISSN: 0730-2312
    Schlagwort(e): neu/p185 protein ; c-erbB-2 ; epidermal growth factor receptor ; transformation ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Growth factor receptors such as the epidermal growth factor receptor (EGFR) and the p185c-neu protein serve vital roles in the transduction of differentiation, developmental, or mitogenic signaling within normal cells. Two methods of analysis suggest that the inappropriately high expression of either protein tyrosine kinase promotes malignant transformation. First, data from in vitro experiments indicate that overexpression of either EGFR or p185c-neu (or the human homolog c-erbB-2) transforms cell-lines. Second, analysis of primary tumors and tumor cell-lines derived from many epithelial tissues (breast, stomach, ovary, and pancreas) show growth factor receptor gene amplification and elevated protein levels. The physical and functional interaction of p185c-neu and EGFR leads to the formation of a highly active, heterodimeric tyrosine kinase complex which synergistically activates cellular transformation. Anti-receptor antibodies have shown potential utility for the down modulation of these cell-surface proteins and suppression of the malignant phenotype. Design of organic antibody “mimetics” based on the structure of antireceptor antibodies may provide useful therapies and biological reagents to affect growth factor receptor function.
    Zusätzliches Material: 4 Ill.
    Materialart: Digitale Medien
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  • 231
    Digitale Medien
    Digitale Medien
    Springer
    BioMetals 5 (1992), S. 73-80 
    ISSN: 1572-8773
    Schlagwort(e): mercury ; arsenic ; cadmium ; plasmid ; restriction analysis ; curing ; conjugation ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie
    Notizen: Abstract Pseudomonas sp isolated from the Bay of Bengal (Madras coast) contained a single large plasmid (pMR1) of 146 kb. Plasmid curing was not successful with mitomycin C, sodium dodecyl sulfate, acridine orange, nalidixic acid or heat. Transfer of mercury resistance from marinePseudomonas toEscherichia coli occurred during mixed culture incubation in liquid broth at 10−4 to 10−5 ml−1. However, transconjugants lacked the plasmid pMR1 and lost their ability to resist mercury. Transformation of pMR1 intoE. coli competent cells was successful; however, the efficiency of transformation (1.49×102 Hgr transformants μg−1 pMR1 DNA) was low.E. coli transformants containing the plasmid pMR1 conferred inducible resistance to mercury, arsenic and cadmium compounds similar to the parental strain, but with increased expression. The mercury resistant transformants exhibited mercury volatilization activity. A correlation existed between metal and antibiotic resistance in the plasmid pMR1.
    Materialart: Digitale Medien
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  • 232
    ISSN: 1432-203X
    Schlagwort(e): Sugarcane ; cell suspension ; protoplast ; microprojectile bombardment ; electroporation ; GUS ; bar ; PAT ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Stably transformed callus of a hybrid sugarcane cultivar (Saccharum species hybrid, CP72-1210) was achieved following high velocity microprojectile bombardment of suspension culture cells, and electroporation of protoplasts. A three-day old cell suspension culture (SC88) was bombarded with gold particles coated with pBARGUS plasmid DNA containing the ß-glucuronidase (GUS) reporter gene and the bar selectable gene that confers resistance to the herbicide basta. The pBARGUS plasmid was also electroporated into the protoplasts of another cell line (SCPP). Colonies resistant to basta were recovered from both sources. Stable integration of the bar gene in the resistant cell lines was confirmed by Southern analysis. In addition, phosphinothricin acetyltransf erase (PAT) activity was also demonstrated in the transformed cell lines.
    Materialart: Digitale Medien
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  • 233
    ISSN: 1432-203X
    Schlagwort(e): Grapevine ; Agrobacterium tumefaciens ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Stem pieces and leaf disks of Vitis spp. were cocultured with Agrobacterium tumefaciens strains carrying the UidA (ß-glucuronidase = GUS) gene. The transformation efficiency was highly increased by using a modified T-6b gene (a gene from pTiTm4) which interferes with normal growth and allows regeneration of normal Nicotiana rustica plants (Tinland 1990). The strains first tested on stem segments were subsequently tested in a leaf explant system. On leaves the transformation efficiency of the strains was much lower than with stems. Both the T-6b gene and the hsp 70-T-6b gene (a modified T-6b gene under the control of a heat shock promoter) allowed the initiation of GUS-positive buds.
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  • 234
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell reports 11 (1992), S. 334-338 
    ISSN: 1432-203X
    Schlagwort(e): Brassica oleracea ; rapid cycling cabbage ; transformation ; Agrobacterium rhizogenes
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Genetically transformed cabbage (Brassica oleracea var. capitata) roots were obtained after inoculation with two engineered Agrobacterium rhizogenes strains, each harbouring a plant selectable marker gene in their T-DNA. Axenic root clones resistant to kanamycin or hygromycin B were established, most of which did not exhibit the phenotypic characteristics of Ri-transformed roots. Shoot regeneration was induced from roots after treatment with 2,4-dichlorophenoxyacetic acid (2,4-D). The resulting plants exhibited various phenotypes: some looked normal, while others showed the transformed phenotype observed in other species. Direct evidence for genetic transformation was obtained by molecular hybridization. The trait was transmitted to the progeny. Transformed cabbage plants can be obtained within 6 months using this approach.
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  • 235
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 19 (1992), S. 715-723 
    ISSN: 1573-5028
    Schlagwort(e): DNase I footprinting ; β-glucuronidase (GUS) ; H-DNA ; transformation ; triple helix
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract There is a 36 bp tract of extreme homopurine/homopyrimidine (PuPy) asymmetry in the maize Adh1 gene promoter (from −44 to −79) that is S1-hypersensitive in plasmids under supercoil tension. Oligodeoxynucleotides corresponding to the PuPy tract were designed to examine the secondary structure of the region and address the possible role of the tract in gene regulation. On the basis of oligodeoxynucleotide band-shift and DNase I footprinting analyses, it was concluded that the homopyrimidine oligodeoxynucleotide can form a triple helix with the duplex PuPy tract in vitro. Transient assays in protoplasts, suspension cells, and seedling roots show that the homopyrimidine oligodeoxynucleotide is also capable of repressing Adh1-GUS gene expression during co-transformation, presumably by the formation of a triple helix with the PuPy tract in vivo. The complementary homopurine oligodeoxynucleotide would not form a triple helix in vitro, nor would it repress Adh1-GUS in vivo. We propose that triple helix formation is a potential regulatory phenomenon in vivo, and that an intraregion triple helix could occur within the Adh1 promoter via the formation of H-DNA.
    Materialart: Digitale Medien
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  • 236
    ISSN: 1573-5028
    Schlagwort(e): dihydrodipicolinate synthase ; lysine overproduction ; potato ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The essential amino acid lysine is synthesized in higher plants by a complex pathway that is predominantly regulated by feedback inhibition of two enzymes, namely aspartate kinase (AK) and dihydrodipicolinate synthase (DHPS). Although DHPS is thought to play a major role in this regulation, the relative importance of AK is not known. In order to study this regulation, we have expressed in the chloroplasts of transgenic potato plants a DHPS derived from Escherichia coli at a level 50-fold above the endogenous DHPS. The bacterial enzyme is much less sensitive to lysine inhibition than its potato counterpart. DHPS activity in leaves, roots and tubers of the transgenic plants was considerably higher and more resistant to lysine inhibition than in control untransformed plants. Furthermore, this activity was accompanied by a significant increase in level of free lysine in all three tissues. Yet, the extent of lysine overproduction in potato leaves was significantly lower than that previously reported in leaves of transgenic plants expressing the same bacterial enzyme, suggesting that in potato, AK may also play a major regulatory role in lysine biosynthesis. Indeed, the elevated level of free lysine in the transgenic potato plants was shown to inhibit the lysine-sensitive AK activity in vivo. Our results support previous reports showing that DHPS is the major rate-limiting enzyme for lysine synthesis in higher plants, but they suggest that additional plant-specific regulatory factors are also involved.
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  • 237
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium ; rice ; transformation ; Ti plasmid ; GUS
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Transient expression of GUS in rice (Oryza sativa L.) mediated by Agrobacterium tumefaciens was characterized using binary vectors containing gusA genes that express minimal (pKIWI105 and pCNL1) or no (p35S-GUS-INT and pCNL56) GUS activity in bacteria. Four-day old seedlings obtained from seeds or immature embryos of rice were cut into shoot, root, and seed remnants and inoculated with various strains of A. tumefaciens. Transient GUS expression events were quantitated histochemically by determining the frequency of explants exhibiting blue spots indicative of GUS at four to six days after cocultivation with A. tumefaciens. A. tumefaciens strains that did not contain the gusA gene (At643) or a Ti-plasmid (At563 and At657) did not elicit any blue staining characteristic of GUS activity. Several parameters were important in obtaining efficient transient expression of GUS in rice mediated by A. tumefaciens. The growth regulator 2,4-D inhibited GUS expression if present during the seed germination period, but the presence of 6 mg/1 2,4-D during cocultivation of the explants with A. tumefaciens slightly enhanced GUS expression efficiency. All 21 rice cultivars tested expressed GUS after co-cultivation with A. tumefaciens. The GUS expression frequency was highest amongst the indica cultivars. The frequencies of GUS expression in japonica cultivars and in Oryza glaberrima cultivars (grown primarily in Africa) were generally one-half to one-third the level found for indica varieties. Leaf explants were more susceptible to A. tumefaciens-facilitated GUS expression than were roots or seed remnants. The vir genes of an agropine-type Ti-plasmid of A. tumefaciens were most effective in directing transient GUS expression in rice, whereas those of a nopaline-type and an octopine-type plasmid were less effective. We have also found that the frequency of transient expression of GUS was higher with pBIN19 as the precursor cloning vector than with pEND4K as the precursor cloning vector. Reasons for differences in effectiveness of these binary vectors are discussed. Using the conditions described here, A. tumefaciens-mediated frequencies of transient GUS expression in four-day old shoots of several rice cultivars were routinely in excess of 50%.
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  • 238
    ISSN: 1573-5028
    Schlagwort(e): hygromycin ; inheritance ; maize ; tissue culture ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Embryogenic maize (Zea mays L.) callus cultures were transformed by microprojectile bombardment with a chimeric hygromycin phosphotransferase (HPT) gene and three transformed lines were obtained by selecting for hygromycin resistance. All lines contained one or a few copies of the intact HPT coding sequence. Fertile, transgenic plants were regenerated and the transmission of the chimeric gene was demonstrated through two complete generations. One line inherited the gene in the manner expected for a single, dominant locus, whereas two did not.
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  • 239
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 18 (1992), S. 201-210 
    ISSN: 1573-5028
    Schlagwort(e): maize ; transformation ; inheritance ; phosphinothricin acetyltransferase ; cotransformation ; microprojectile bombardment
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Progeny recovered from backcrossed transgenic maize tissue culture regenerants (R0) were analyzed to determine the segregation, expression, and stability of the introduced genes. Transgenic A188×B73 R0 plants (regenerated from embryogenic suspension culture cells transformed by microprojectile bombardment; see [9]) were pollinated with nontransformed B73 pollen. Inheritance of a selectable marker gene, bar, and a nonselectable marker gene, uidA, was analyzed in progeny (R1) representing four independent transformation events. Activity of the bar gene product, phosphinothricin acetyltransferase (PAT), was assessed in plants comprising the four R1 populations. The number of R1 plants containing PAT activity per total number of R1 plants recovered for each population was 2/7, 19/34, 3/14 and 73/73. Molecular analysis confirmed the segregation of bar in three R1 populations and the lack of segregation in one R1 population. Cosegregation analysis indicated genetic linkage of bar and uidA in all four R1 populations. Analysis of numerous R2 plants derived from crossing transformed R1 plants with nontransformed inbreds revealed 1:1 segregation of PAT activity in three of four lines, including the line that failed to segregate in the R1 generation. Integrated copies of bar in one line appeared to be unstable or poorly transmitted.
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  • 240
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 18 (1992), S. 835-839 
    ISSN: 1573-5028
    Schlagwort(e): transformation ; particle gun ; soybean ; GUS gene
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We observed that flowing helium at moderate pressures accelerated DNA-coated microprojectiles to velocities suitable for penetration of cells in intact plant tissues. The flowing helium principle permitted the construction of a simple and inexpensive transformation device that was easier to use than those previously described. This device provided efficient transformation of cells in soybean seedlings and other plants.
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  • 241
    ISSN: 1573-5028
    Schlagwort(e): patatin class II gene ; Solanum tuberosum ; transformation ; transgenic potato ; tuber-specific gene expression
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract From a potato genomic library a phage lambda clone was isolated that carried nucleotide sequences of two patatin genes, thus demonstrating a close physical linkage between these two members of the patatin gene family. Sequence and restriction analysis showed the genes to be oriented in tandem. The more upstream gene was a pseudogene truncated at the 3′ end, whereas the downstream gene was a class II patatin gene. In addition to a 208 bp fragment also present in patatin class I promoters, the region in between both genes contained various direct repeats also found in other patatin genes. To study the promoter activity of this intergenic region, a 2.78 kb fragment was transcriptionally fused to the β-glucuronidase gene and reintroduced into potato cultivar Bintje. Histochemical analysis revealed expression in the outermost layer of cells of the cortex, in the tuber phellogen, in or around the root vascular system, and also in the abaxial phloem layer of the vascular bundle in leaves.
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  • 242
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium tumefaciens ; β-glucuronidase ; meristem ; microprojectile bombardment ; neomycin phosphotransferase ; sunflower ; tobacco ; transformation ; wounding
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Bombardment of plant tissues with microprojectiles in an effective method of wounding to promote Agrobacterium-mediated transformation. Tobacco cv. Xanthi leaves and sunflower apical meristems were wounded by microprojectile bombardment prior to application of Agrobacterium tumefaciens strains containing genes within the T-DNA encoding GUS or NPTII. Stable kanamycin-resistant tobacco transformants were obtained using an NPTII construct from particle/plasmid, particle-wounded/Agrobacterium-treated or scalpel-wounded/Agrobacterium-treated potato leaves. Those leaves bombarded with particles suspended in TE buffer prior to Agrobacterium treatment produced at least 100 times more kanamycin-resistant colonies than leaves treated by the standard particle gun transformation protocol. In addition, large sectors of GUS expression, indicative of meristem cell transformation, were observed in plants recovered from sunflower apical explants only when the meristems were wounded first by particle bombardment prior to Agrobacterium treatment. Similar results in two different tissue types suggest that (1) particles may be used as a wounding mechanism to enhance Agrobacterium transformation frequencies, and (2) Agrobacterium mediation of stable transformation is more efficient than the analogous particle/plasmid protocol.
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  • 243
    ISSN: 1573-5028
    Schlagwort(e): coat protein ; potato virus Y ; tobacco ; transformation ; resistance
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The coat protein (CP) cistron of the tobacco veinal necrosis strain of potato virus Y (PVYN), supplemented with translational start signals, was cloned into an Agrobacterium tumefaciens Ti transformation vector. Transformation of tobacco leaf discs resulted in 99 transgenic lines which were subsequently analysed for the presence and expression, at both the transcriptional and translational level, of the CP-gene. Although CP-specific RNA transcripts were produced in all plants no CP could be detected by several sensitive immunological techniques. Upon mechanical inoculation of progeny lines of selfpollinated original transformants (S1) with PVYN, protection levels of 20 and 95%, respectively, could be observed in two out of ten lines tested. This level of protection increased to 100% in the S2 progeny obtained from self-pollination of virus-protected S1 plants. Transformation of tobacco leaf discs with a PVYN CP construct from which the ATG start codon had been removed by site-directed mutagenesis resulted in 57 transgenic lines that all produced CP-specific transcripts. Mechanical inoculation with PVYN of S1 progeny plants of several of these lines resulted in resistance to a similar level and extent as in the S1 progeny of plants transformed with the intact CP cistron. The results obtained strongly suggest that the resistance observed in the transgenic plants is principally based on the presence of PVYN CP RNA sequences rather than on the accumulation of viral coat protein.
    Materialart: Digitale Medien
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  • 244
    ISSN: 1432-203X
    Schlagwort(e): Agrobacterium tumefaciens ; Solanum integrifolium ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary The wild species Solanum integrifolium represents a source of pest and disease resistance genes for breeding strategies of the cultivated species Solanum melongena. Somatic hybridization via protoplast fusion between the two species may provide a valuable tool for transferring polygenic traits into the cultivated species. The availability of S.integrifolium cells carrying dominant selectable markers would facilitate the heterokaryon rescue. An appropriate methodology for in vitro culture and plant regeneration from leaf explants of S.integrifolium is reported. Efficient leaf-disk transformation via co-cultivation with Agrobacterium tumefaciens led to the regeneration of transformed plants carrying the reporter genes GUS and NPT-II. Transformed individuals were obtained through selection on kanamycin-containing medium. Stable genetic transformation was assessed by histochemical and enzymatic assays for GUS and NPT-II activity, by the ability of leaf disks to initiate callus on Km-containing medium, Southern blot analyses of the regenerated plants, and genetic analysis of their progenies. Selfed-seed progeny of individual transformed plants segregated seedlings capable to root and grow in selective condition, while untransformed progeny did not. Genetic analyses of progeny behaviour showed that the reporter gene NPT-II segregated as single as well as two independent Mendelian factors. In two cases an excess of kanamycin-sensitive seedlings was obtained, not fitting into any genetic hypothesis.
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  • 245
    Digitale Medien
    Digitale Medien
    Springer
    Russian chemical bulletin 41 (1992), S. 2119-2120 
    ISSN: 1573-9171
    Schlagwort(e): transformation ; thiocyanoalkyl phosphites ; amidophosphites
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Chemie und Pharmazie
    Notizen: Abstract The ratio of the alkoxy and dialkylamido groups in thiocyanoalkyl phosphites determines the structure of the transformation products of these compounds.
    Materialart: Digitale Medien
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  • 246
    ISSN: 1573-6881
    Schlagwort(e): Mitochondria ; hexokinase ; normal and tumor cells ; enzyme localization ; subcellular fractionation ; receptor for binding ; monoamine oxidase ; NADPH-cytochromec reductase ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Physik
    Notizen: Abstract Hexokinase plays an important role in normal glucose-utilizing tissues like brain and kidney, and an even more important role in highly malignant cancer cells where it is markedly overexpressed. In both cell types, normal and transformed, a significant portion of the total hexokinase activity is bound to particulate material that sediments upon differential centrifugation with the crude “mitochondrial” fraction. In the case of brain, particulate binding may constitute most of the total hexokinase activity of the cell, and in highly malignant tumor cells as much as 80 percent of the total. When a variety of techniques are rigorously applied to better define the particulate location of hexokinase within the crude “mitochondrial fraction,” a striking difference is observed between the distribution of hexokinase in normal and transformed cells. Significantly, particulate hexokinase found in rat brain, kidney, or liver consistently distributes with nonmitochondrial membrane markers whereas the particulate hexokinase of highly glycolytic hepatoma cells distributes with outer mitochondrial membrane markers. These studies indicate that within normal tissues hexokinase binds preferentially to non-mitochondrial receptor sites but upon transformation of such cells to yield poorly differentiated, highly malignant tumors, the overexpressed enzyme binds preferentially to outer mitochondrial membrane receptors. These studies, taken together with the well-known observation that, once solubilized, the particulate hexokinase from a normal tissue can bind to isolated mitochondria, are consistent with the presence in normal tissues of at least two different types of particulate receptors for hexokinase with different subcellular locations. A model which explains this unique transformation-dependent shift in the intracellular location of hexokinase is proposed.
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  • 247
    Digitale Medien
    Digitale Medien
    Springer
    World journal of microbiology and biotechnology 8 (1992), S. 92-97 
    ISSN: 1573-0972
    Schlagwort(e): Bacteriocin ; conjugation ; electroporation ; Lactobacillus acidophilus ; protoplast fusion ; plasmids ; transduction ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Werkstoffwissenschaften, Fertigungsverfahren, Fertigung
    Notizen: Abstract Lactobacillus acidophilus has been recommended as a dietary adjunct because of its antagonistic action toward intestinal pathogens and anti-carcinogenic and hypocholesterolemic activities. ManyL. acidophilus strains harbour plasmids and such strains generally produce bacteriocin(s). Resistance to antibiotics has also been shown to be linked with plasmids. Gene transfer and cloning systems are being developed forL. acidophilus which should permit the rapid genetic characterization of desired species and their modification to obtain predetermined traits. Drug resistance determinants and production of antibiotic-like substances may serve as suitable markers for the study and development of these genetic systems. Recent developments in gene transfer systems have been reviewed here.
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  • 248
    Digitale Medien
    Digitale Medien
    Springer
    Nutrient cycling in agroecosystems 32 (1992), S. 313-319 
    ISSN: 1573-0867
    Schlagwort(e): Maize-mustard crop sequence ; ‘methanised’ FYM-bioslurry ; inter-relationships ; zinc fractions ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Organic matter is the major source of zinc in soil. The availability of this nutrient is dependent on the release from organic matter through mineralization and reaction with soil particles. Addition of bioslurry, containing 70 mg Zn kg−1 will influence availability of Zn through its effect on transformation reaction in soil. The present study was conducted to determine the distribution of major chemical forms of Zn in an alluvial soil, to understand the changes in zinc fractions due to bioslurry application and cropping and to find out the inter-relationships and equilibria between the fractions. Soil solution + exchangeable Zn (Zn-CA), specifically sorbed Zn by inorganic sites (Zn-ACC), specifically sorbed Zn by organic sites (Zn-PYR), Zn occluded by free oxides (Zn-OX), and residual zinc (Zn-RES) constituted 0.3, 4.5, 16.6, 16.3 and 57.3 percent, respectively of the total Zn content (Zn-TOT). Application of 13.32 t ha−1 bioslurry increased Zn content in Zn-CA, Zn-PYR and Zn-RES by 72.7, 93.2 and 36.4 percent, respectively over control. Zn occluded by free oxides (Zn-OX) was found released by the dissolution action of organic compounds present in bioslurry and the amount of Zn so released was transformed to Zn-RES, Zn-CA and Zn-DTPA. Growing crops increased Zn content in Zn-RES fraction only. Linear positive relationships between Zn-CA, Zn-PYR, Zn-RES and DTPA-Zn and bioslurry levels marked the significance of bioslurry in stabilising the status of these fractions. Path coefficient analysis and intercorrelation studies indicated the existence of equilibrium between different Zn fractions in soils.
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  • 249
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 30 (1992), S. 141-148 
    ISSN: 1573-5044
    Schlagwort(e): electroporation ; pea ; Pisum sativum L. ; protoplast ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Protoplasts from two different pea cultivars, Belman and Filby, were stably transformed by direct gene transfer using electroporation. Transgenic calli could be obtained after selection, when hygromycin resistance was used as the selective trait introduced into the protoplasts, while no transformants were obtained when kanamycin resistance was used as selective marker in either of the two pea cultivars tested. The effect of the field strength on survival and division rates of the protoplasts was studied. Two different culture systems and osmotica were compared for induction of sustained divisions in and regeneration of transgenic callus from the protoplasts. The choice of the culture system had a considerable effect on the initial division frequency of the treated protoplasts, as well as on the later growth of the colonies. Transformation efficiency was monitored by histochemical GUS assay, and the transgenic nature of the calli selected for resistance against antibiotics was confirmed by DNA analysis.
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  • 250
    Digitale Medien
    Digitale Medien
    Springer
    Applied mathematics and mechanics 13 (1992), S. 1149-1162 
    ISSN: 1573-2754
    Schlagwort(e): plate ; displacement ; nonlinear ; transformation ; perturbation method
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Maschinenbau , Mathematik , Physik
    Notizen: Abstract To begin with, in this paper, the displacement governing equations and the boundary conditions of nonsymmetrical large deflection problem of circular thin plates are derived. By using the transformation and the perturbation method, the nonlinear displacement equations are linearized, and the approximate boundary value problems are obtained. As an example, the nonlinear bending problem of circular thin plates subjected to comparatively complex loads is studied.
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  • 251
    Digitale Medien
    Digitale Medien
    Springer
    Cytotechnology 10 (1992), S. 93-124 
    ISSN: 1573-0778
    Schlagwort(e): hybridomas ; embryonic stem cells ; immortalization ; amine oxidase ; polyamines ; cell death ; crisis ; transformation ; aneuploidy ; senescence
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin , Werkstoffwissenschaften, Fertigungsverfahren, Fertigung
    Notizen: Abstract It has been known for several decades that cultured murine cells undergo a defined series of changes, i.e., anin vitro evolution, which includes crisis, spontaneous transformation (‘immortalization’), aneuploidy, and spontaneous neoplastic transformation. These changes have been shown to be caused by thein vitro environment rather than an inherent instability of the murine phenotype or genotype. Serum amine oxidases were recently identified as a predominant cause of crisis. These enzymes generate hydrogen peroxide from polyamine substrates that enter the extracellular milieu. This finding implicates free-radical toxicity as the underlying cause ofin vitro evolution. We propose an oxyradical hypothesis to explain each of the stages ofin vitro evolution and discuss its significance for cytotechnology and long-term cultivation of mammalian cell types.
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  • 252
    Digitale Medien
    Digitale Medien
    Springer
    Bulletin of experimental biology and medicine 113 (1992), S. 532-535 
    ISSN: 1573-8221
    Schlagwort(e): stromal cells ; transformation ; nude mice
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Materialart: Digitale Medien
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  • 253
    ISSN: 0749-503X
    Schlagwort(e): Candida maltosa ; electroporation ; transformation ; plasmid vectors ; Life and Medical Sciences ; Genetics
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: Conditions for efficient and quick transformation by electroporation were developed in Candida maltosa. To investigate the efficiency of transformation with integrative as well as with autonomously replicating plasmids, a series of vectors was constructed for homologous transformation of the species. Transformants were obtained with different plasmids as covalently closed circular molecules and as linearized DNA. The influence of recipient strain and plasmid type as well as of cell number and parameters of the supplied electrical pulse on the transformation efficiency have been investigated. A maximum of 7000 transformants per 100ng of plasmid DNA was reached. The efficiency of transformation was compared with that of the LiCl method.
    Zusätzliches Material: 5 Ill.
    Materialart: Digitale Medien
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  • 254
    ISSN: 0749-503X
    Schlagwort(e): Candida maltosa ; automonous replicating sequence ; nucleotide sequence ; transformation ; RS15 protein ; intron ; Life and Medical Sciences ; Genetics
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: A Candida maltosa chromosomal DNA fragment which confers high frequency transformation of C. maltosa and autonomous replication of recombinant plasmids was cloned and sequenced. Analysis of the nucleotide sequence of the cloned DNA revealed a sequence homologous for C. maltosa autonomously replicating sequence (ARS) elements. Vector pRJ1 for C. maltosa was constructed, which contained a 1.3 kb ARS sequence, pICEM-19H and the ADE1 gene of C. maltosa. Southern blot analysis suggested that the copy number of pRJ1 in C. maltosa was approximately 20 per genome. The sequence analysis also revealed an open reading frame, encoding a polypeptide with high homology (70%) to the RS15 protein of Brugia pagangi. This open reading frame has an intron with canonical sites for correct splicing in Saccharomyces cerevisiae.
    Zusätzliches Material: 5 Ill.
    Materialart: Digitale Medien
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  • 255
    Digitale Medien
    Digitale Medien
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 13 (1992), S. 118-125 
    ISSN: 0192-253X
    Schlagwort(e): Microinjection ; macronucleoplasm ; transformation ; Paramecium ; Life and Medical Sciences ; Genetics
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: Transformation by microinjection of macronucleoplasm in Paramecium caudatum was investigated. Macronucleoplasm with three genetic markers (behavior, trichocyst, and mating type) was injected into the macronucleus. To facilitate microinjection, in most cases, paramecia were immobilized in a gelatin (7.5%) solution. The injected cells began to express a dominant gene (cnrA+ or cnrB+) of the donor 9-24 hr after injection. Expression did not require cell division suggesting injected macronucleoplasm was capable of expressing a phenotype. The amount of injected macronucleoplasm appears to correlate with the frequency of successful expression but not to correlate with the time required for expression. After a number of fissions, the injected cells produced clones which had cells expressing the phenotype of the donor. This suggests that injected macronucleoplasm was replicated and expressed in the recipient cell lines. The transformed clones were classified into two groups. In one group, transformation was stable. All cell lines derived from the injected cells expressed a phenotype similar to the heterozygote of donor and recipient cells. In the other group, transformation was unstable. During the first five to seven fissions after injection, at each division, cells produced one daughter cell which later reverted to the recipient phenotype. After this unstable period, cells no longer produced the recipient phenotype but produced the donor phenotype exclusively. Donor and recipient phenotypes were, thus, segregated in different cell lines. Observation of genetic markers and analysis by computer simulation shed light on the mode of transmission of injected macronucleoplasm. In stable transformation, injected macronucleoplasm appears to be distributed equally to daughter cells. In unstable transformation, injected macronucleoplasm is distributed only to one of the daughter cells at every division until about the fifth to seventh fission after injection and then begins to assort equally to daughter cells. The cell cycle stage at injection may influence the mode of transformation. Interspecific microinjection of macronucleoplasm from P. multimicronucleatum and P. tetraurelia to P. caudatum. resulted in the expression of foreign genes in P. caudatum. In one case, injection of macronucleoplasm of P. tetraurelia produced a stable transformant indicating replication of foreign macronucleoplasm in P. caudatum. This work reveals the mode of transformation by injected macronucleoplasm and shows the possibility of transformation among Paramecium species, which is significant in the study of the conservation of gene products and the mechanism of gene expression in different species. © 1992 Wiley-Liss, Inc.
    Zusätzliches Material: 2 Ill.
    Materialart: Digitale Medien
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  • 256
    ISSN: 1432-0428
    Schlagwort(e): Islet cell antibodies ; Type 1 (insulin-dependent) diabetes ; Epstein-Barr virus ; peripheral blood lymphocytes ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Islet cell antibodies are usually detected in the sera of almost all Type 1 (insulin-dependent) diabetic patients within several months after onset of the disease. The antibodies then disappear quite early during the course of the disease. The present study was undertaken to detect islet cell antibody-producing clones in peripheral blood lymphocytes of Type 1 diabetic patients whose islet cell antibodies could not be detected in sera. Epstein-Barr virus-transformed lymphocytes were employed to enhance the production of antibodies and to detect the clones from peripheral blood lymphocytes. Peripheral blood lymphocytes were obtained from 40 islet cell antibody-negative Type 1 diabetic patients, 10 antibody-positive Type 1 diabetic patients, 30 Type 2 (non-insulin-dependent) diabetic patients and 40 normal control subjects. Epstein-Barr virus-transformed lymphocytes were cultured for 4 weeks and the culture supernatants were used for assay of islet cell antibodies. Islet cell antibody assays were performed by immunohistochemical methods using peroxidase-labelled protein A for IgG antibodies, peroxidase-labelled anti-human IgM antibodies for IgM antibodies and fresh frozen human pancreatic tissue. IgG-islet cell antibodies were detected in 26 islet cell antibody-negative patients (65%), eight antibody-positive patients (80%) and one Type 2 diabetic patient (3%) in the culture supernatants. Islet cell antibodies in the supernatants could not be detected in any of the control subjects. IgM-islet cell antibodies could not be detected in any of the patients or control subjects. These findings indicate that islet cell antibody-producing clones exist in peripheral blood lymphocytes from Type 1 diabetic patients whose islet cell antibodies cannot be detected in their sera and IgG-islet cell antibodies might be a specific characteristic of Type 1 diabetes. The detection of islet cell antibodies from Epstein-Barr virus-transformed lymphocytes may be useful in examining the role of autoimmune mechanisms in the development of disease.
    Materialart: Digitale Medien
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  • 257
    ISSN: 1432-203X
    Schlagwort(e): Agrobacterium tumefaciens ; Allium cepa ; Antirrhinum majus ; Brassica campestris ; Glycine max ; Nicotiana tabacum ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Expiants of five plant species (Allium cepa, Antirrhinum majus, Brassica campestris. Glycine max, and Nicotiana tabacum) were co-cultivated with three Agrobacterium tumefaciens strains under different conditions to assess the effects of acetosyringone and medium pH on strain virulence. Tumours were incited on all dicotyledonous species by strains N2/73 and A281. The presence of acetosyringone during co-cultivation generally enhanced the virulence of these strains, most markedly N2/73 on A. majus and G. max, and A281 on G. max. Strain Ach5 was virulent only on N. tabacum in the absence of acetosyringone, which, when present, extended the host range to include A. majus. There was evidence to suggest that acetosyringone may suppress virulence in some strain/plant species interactions. Virulence was affected in some cases by medium pH, but there was no general effect across plant species.
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  • 258
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell reports 10 (1991), S. 85-89 
    ISSN: 1432-203X
    Schlagwort(e): Agrobacterium rhizogenes ; alkamides ; Echinacea purpureal ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Echinacea purpurea seedlings were inoculated with several Agrobacterium rhizogenes strains in order to obtain hairy roots. Infection with A. rhizogenes strains LMG63 and LMG150 resulted in callus formation. Upon infection with strains ATCC 15834 and R1601 hairy roots were obtained. Opine detection confirmed transformation of E. purpurea. Comparative HPLC fingerprint analysis of the alkamides from natural plant source, control tissues, and transformed callus and roots indicated that transformed callus and hairy roots might be a promising source for continuous and standardized production of the dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamide and related amides.
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  • 259
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 105 (1991), S. 171-177 
    ISSN: 1573-4919
    Schlagwort(e): estrogen receptor ; transformation ; aging ; rat uterus
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract The conformation of estrogen receptor (ER) and its in vitro transformation by RNase, Urea and ATP were analysed using the uteri of young (16 weeks) and old (92 weeks) rats. Following the digestion of ER with proteolytic enzymes like trypsin and chymotrypsin and the analysis of cleaved fragments by SDS-PAGE, similar pattern is observed in both ages. In vitro transformation of ER by RNase, Urea and ATP shows that the degree of transformation is lower in old than young. Furthermore, the transformed ER from old is less capable of binding to DNA than that from young. Thus our results show that the conformation of ER probably does not change with age, but the degree of transformation and the ability of transformed receptor to bind to DNA decrease with age.
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  • 260
    ISSN: 1573-5028
    Schlagwort(e): methylation ; Oryza sativa ; protoplasts ; transformation ; β-D-glucuronidase ; methotrexate ; hygromycin
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Cell suspension-derived rice (Oryza sativa L.) protoplasts were transformed by direct gene uptake. PEG-mediated transformation was more efficient than electroporation. Plasmid DNA containing a hygromycin phosphotransferase (HPT) gene (which confers hygromycin resistance) driven by the CaMV 35S promoter and a β-D-glucuronidase (GUS) gene under control of the 1′, 2′ double promoter of the mannopine synthase (mas) locus of Agrobacterium tumefaciens was introduced into rice protoplasts. Southern analysis of DNA from transformed cell lines showed that the HPT and GUS genes were present intact. Both genes were expressed in transgenic cell suspensions. GUS activity was detected by histochemical staining of the cells and by enzyme assays. During a 12-day culture period the proportion of stained cells rose to a maximum and then decreased again. Considerably higher numbers of blue-stained cells were obtained when the transgenic cell lines were grown in the presence of 5-azacytidine. Transcripts of the GUS gene could not be detected, in contrast with the HPT gene. Plantlets were regenerated from one transgenic cell line. GUS activity was found in both leaf and root tissues of these plants, particularly, but not exclusively, in vascular bundles. A mouse dihydrofolate reductase coding sequence (DHFR), conferring methotrexate resistance, fused to the CaMV 35S promotor and the wild-type nopaline synthase (NOS) gene of A. tumefaciens were also introduced into rice protoplasts. Stable integration of both genes was confirmed by Southern analysis. Expression of the DHFR gene was demonstrated by high levels of resistance to methotrexate of the transgenic cell suspensions and by the presence of DHFR transcripts. Expression of the NOS gene at enzyme or RNA level was not detected. Southern analysis suggests that this gene was probably either methylated or scrambled in these lines.
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  • 261
    ISSN: 1573-4943
    Schlagwort(e): Conformational energy ; three-dimensional structure ; amino acid substitution ; c-abl oncogene ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Chemie und Pharmazie
    Notizen: Abstract Thebcr-abl chimeric gene of Philadelphia chromosome positive chronic myelogenous leukemias is only weakly transforming. This transformation activity is greatly enhanced by a Lys-for-Glu substitution at position 832 in the c-abl gene, as occurs in the highly transforming v-abl genes. It has been suggested that this mutation results in a significant structural change in the encoded protein product. Using conformational energy analysis, we have determined the allowed low-energy conformations for residues 828–836 of this protein with Lys and Glu at position 832. In both cases, the overwhelmingly preferred conformation for this region is a bend-helix motif. The helix terminates at residue 836, and there are no discernible differences in conformation between the Lys- and Glu-containing sequences. These results suggest that the activating amino acid substitution at position 832 in the c-abl protein product does not produce its effect via a local conformational change.
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  • 262
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 16 (1991), S. 263-269 
    ISSN: 1573-5028
    Schlagwort(e): transformation ; enhancer trap ; β-glucuronidase ; potato
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A vector has been designed that contains a truncated CaMV (cauliflower mosaic virus) 35S promoter fused to a receptor gene encoding β-glucuronidase (GUS), placed adjacent to the left border sequence of an Agrobacterium vector. In potato plants transformed with this vector, different patterns of transcription were detected at high frequency using in situ assays for GUS activity. Previous studies in Drosophila using analogous vectors have shown that the new patterns of transcription in many cases reflect the patterns of expression of genes adjacent to the site of vector insertion. If this is also the case in plants, the vector described here will be useful in identifying the activity of genes in different cell types and will assist in determining their function.
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  • 263
    ISSN: 1573-5028
    Schlagwort(e): recombinant ; phosphoenolpyruvate carboxylase ; C4 plant ; cDNA ; transformation ; Escherichia coli ; protein phosphorylation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Phosphoenolpyruvate carboxylase (PEPC)-deficient mutants ofEscherichia coli have been complemented with a plasmid bearing a full-length cDNA encoding the C4-type form ofSorghum leaf PEPC. Transformed cells grew on minimal medium. Two clones were selected which produce a functional and full-sized enzyme protein as determined by activity assays, immunochemical behavior and SDS-PAGE. In addition, regulatory phosphorylation of immunopurified recombinant PEPC was observed when the enzyme was incubated with a partially purified plant PEPC kinase. These results establish thatE. coli cells produce a genuine, phosphate-free, higher-plant PEPC. Application of immunoadsorbtion chromatography to bacterial extracts makes it possible to prepare highly pure protein available for biochemical studies.
    Materialart: Digitale Medien
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  • 264
    ISSN: 1432-1424
    Schlagwort(e): intercellular communication ; gap junction ; connexin ; growth control ; cDNA ; connexin43 ; cell-cell channel ; junctional communication ; transformation ; cancer etiology
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie
    Notizen: Summary Incorporation of the gene for connexin 43, a cell-cell channel protein of gap junction, into the genome of communication-deficient transformed mouse 10T1/2 cells restored junctional communication and inhibited growth. Growth was slowed, saturation density reduced and focus formation suppressed, and these effects were contingent on overexpression of the exogenous gene and the consequent enhancement of communication. In coculture with normal cells the growth of the connexin overexpressors was completely arrested, as these cells established strong communication with the normal ones. Thus, in culture by themselves or in coculture, the connexin overexpressor cells grew like normal cells. These results demonstrate that the cell-cell channel is instrumental in growth control; they are the expected behavior if the channel transmits cytoplasmic growth-regulatory signals.
    Materialart: Digitale Medien
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  • 265
    Digitale Medien
    Digitale Medien
    Springer
    Cancer and metastasis reviews 10 (1991), S. 141-150 
    ISSN: 1573-7233
    Schlagwort(e): melanocyte ; melanoma ; differentiation ; transformation ; antigen
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Melanoma is a valuable model to study phenotypic traits that are regulated during cell differentiation and malignant transformation. Melanoma cells display extensive phenotypic and antigenic heterogeneity. Studies of this attribute have provided insight into events that take place during normal melanocyte differentiation and give clues to traits that contribute to malignancy. It is possible that the phenotypic and genotypic heterogeneity present among melanoma cells within a single lesion includes a subset of cells with traits that favor tumor progression and metastasis. This review discusses the identification and characterization of antigens expressed by melanoma cells and their potential contribution to melanocyte differentiation and malignant transformation.
    Materialart: Digitale Medien
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  • 266
    ISSN: 1573-5036
    Schlagwort(e): blue-green algae ; iron ; manganese ; submerged soils ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract N2-fixing blue-green algae (Cyanobacteria), besides enriching soils with N and organic carbon, may modify a number of chemical and electro-chemical properties of the soils resulting in a change in availability of some micronutrient elements. Keeping this in view, an experiment was conducted to study the effects of growth and subsequent decomposition of blue-green algae on changes in the different forms of Fe and Mn in four soils under submerged condition. A mixed algal culture containing Anabaena, Nostoc, Cylindrospermum, and Tolypothrix was used as inoculum. It was allowed to grow for 2 months, after which the soils were sequentially extracted with (i) M NH4OAc (pH 7.0), (ii) M K4P2O7, (iii) 0.1 M NH2OH.HCl (pH 2.0), (iv) 0.2 M (NH4)2C2O4 (pH 3.0) and (v) 0.1 M ascorbic acid to obtain water-soluble plus exchangeable, organically bound, easily reducible, amorphous oxides-and crystalline oxides-bound forms of Fe and Mn, respectively, both during the growth as well as the subsequent in-situ decomposition of the algal biomass in soils. Iron and Mn in the extracts were estimated by atomic absorption spectrophotometry. The results showed that growth of blue-green algae in submerged rice soils caused a decrease in the NH4OAc-extractable forms of Fe and Mn with concomitant increases in all the other four determined forms of the elements. Such decreases and/or increases in different forms of Fe and Mn in soils were explained as being due to release of O2, addition of organic matter and liberation of extracellular organic compounds by the blue-green algae during their growth. The decomposition of algal biomass resulted in an increase in the NH4OAc-, K4P2O7- and (NH4)2C2O4-extractable forms of Fe and Mn with a simultaneous decrease in the NH2OH · HCl- and ascorbic acid-extractable forms. Development of strong reducing conditions and formation of organic acids with chelating properties were suggested as being the cause of the above changes. The implication of these changes in the forms of Fe and Mn for the Fe and Mn nutrition of rice plants were discussed.
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  • 267
    ISSN: 1573-5060
    Schlagwort(e): Agrobacterium rhizogenes ; antisense RNA ; granule-bound starch synthase ; Solanum tuberosum ; starch composition ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content.
    Materialart: Digitale Medien
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  • 268
    Digitale Medien
    Digitale Medien
    Springer
    Euphytica 55 (1991), S. 157-169 
    ISSN: 1573-5060
    Schlagwort(e): gene transfer ; genetic manipulation ; chimaeric genes ; legumes ; transformation ; somatic hybridisation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Summary The merits and limitations of somatic cell techniques involving Agrobacterium-mediated transformation, direct gene transfer and protoplast fusion, are discussed in relation to the genetic improvement of forage and grain legumes. Whilst progress with legumes is limited compared to that with plants of other families such as the Solanaceae, the fact that many legumes are readily amenable to tissue culture now permits somatic cell techniques to be targetted to these species. Future development of the subject will necessitate close collaboration between molecular biologists and plant breeders to enable novel plants generated by in vitro technologies to be incorporated into conventional breeding programmes.
    Materialart: Digitale Medien
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  • 269
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 24 (1991), S. 91-95 
    ISSN: 1573-5044
    Schlagwort(e): Agrobacterium ; regeneration ; Ribes nigrum ; tissue culture ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Transformation of the black currant cv. Ben More was achieved by utilising the binary vector system of Agrobacterium tumefaciens. This system involved the inoculation of peeled internodal stem segments with A. tumefaciens strain LBA4404 containing the binary vector PBI121.X with the marker genes Betaglucuronidase (GUS) and neomycin phosphotransferase II (NPTII). Shoot regeneration occurred on nutrient media based on M&S salts. Transformation was confirmed by the fluorogenic assay procedure which determined that the GUS gene had been transferred into the plant material and was being expressed. Concurrent transfer of the NPTII gene into the plant material was also confirmed with a ‘dot blot’ assay on selected GUS positive plantlets.
    Materialart: Digitale Medien
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  • 270
    Digitale Medien
    Digitale Medien
    Springer
    Journal of bioenergetics and biomembranes 23 (1991), S. 903-917 
    ISSN: 1573-6881
    Schlagwort(e): Transferrin receptor ; diferric transferrin ; 3T3 cells ; transformation ; iron reduction ; plasma membrane
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Physik
    Notizen: Abstract Transformation of 3T3 cells by SV40 virus changes the properties of the transplasma membrane electron transport activity which can be assayed by reduction of external ferric salts. After 42 h of culture and before the growth rate is maximum, the transformed cells have a much slower rate of ferric reduction. The change in activity is expressed both by change inK m andV max for ferricyanide reduction. The change in activity is not based on surface charge effect or on tight coupling to proton release or on intracellular NADH concentration. With transformation by SV40 virus infection the expression of transferrin receptors increases, which correlates with greater diferric transferrin stimulation of the rate of ferric ammonium citrate reduction in transformed SV40-3T3 cells than in 3T3 cells.
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  • 271
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 24 (1991), S. 163-172 
    ISSN: 1573-5044
    Schlagwort(e): Agrobacterium rhizogenes ; cryopreservation ; hairy roots ; molecular stability ; secondary metabolites ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Crypopreservation methods were firstly developed for root-tips from hairy root cultures of Beta vulgaris, established after transformation by Agrobacterium rhizogenes. The effects of culture age, pre-growth, cryoprotection, freezing rate and post-freeze culture conditions were determined. The resulting freezing protocol was then used to cryopreserve transformed root cultures of Nicotiana rustica. Both species were viable after freezing (ca. 80%), according to fluorescein diacetate vital staining. However, on average the regeneration of proliferating roots from surviving root-tips was low (〈20%). Growth rates, secondary metabolite production and T-DNA structure of a number of hairy root lines were examined and found to be unchanged after cryopreservation.
    Materialart: Digitale Medien
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  • 272
    ISSN: 1573-0603
    Schlagwort(e): Epstein-Barr virus ; feeder layer ; B-lymphocytes ; transformation ; human cell lines
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Methods are described for the efficient transformation of fresh or cryopreserved human B lymphocytes to produce continuous, lymphoblastic cell lines. Lymphocytes are separated from whole blood by centrifugation through Ficoll. They are transformed by exposure to Epstein-Barr Virus (EBV) obtained as a supernatant from ATCC. CRL 1612 (B95-8) cells. Virus production is verified in advance by immunoperoxidase staining after application of a monoclonal antibody to EBV capsid antigen [ATCC.HB 168 (72A1)]. The addition of irradiated feeder cells [ATCC.CCL 17 (MRC-5)] is important to enhance efficiency in lymphoblast culture initiation.
    Materialart: Digitale Medien
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  • 273
    Digitale Medien
    Digitale Medien
    Springer
    Journal of automated reasoning 7 (1991), S. 337-358 
    ISSN: 1573-0670
    Schlagwort(e): Control rules ; transformation ; logic programming
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Informatik
    Notizen: Abstract We present a technique for the compilation of bottom-up and mixed logic derivations into PROLOG-programs. It is obtained as an extension of a program transformation technique called Compiling Control. We illustrate its applications in three different domains: solving numerical problems, integrity checking in deductive databases and theorem proving. The aim is to obtain efficient PROLOG programs for problems in which a non-top-down control is most appropriate.
    Materialart: Digitale Medien
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  • 274
    ISSN: 1573-5044
    Schlagwort(e): Agrobacterium ; leaf explant ; mesophyll protoplast ; regeneration ; selective agent ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Agrobacterium tumefaciens strains harbouring plasmid vectors pBCAT1, pVU1011 or pMON806 were used to transform leaf explants of Nicotiana tabacum cultivars ‘Delgold’ and ‘Candel’, N. debneyi, and N. rustica var. NRT. Transgenic plants resistant to the selective agents kanamycin, hygromycin or methotrexate were regenerated and used as sources of leaf mesophyll protoplasts. Protoplasts divided and regenerated plants in the presence of selective agents at levels inhibitory to protoplasts of non-transformed plants. Cross-resistance of protoplasts to more than one selective agent was not observed in this study which suggests that this approach may lead to an efficient interspecific somatic hybrid selection system.
    Materialart: Digitale Medien
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  • 275
    Digitale Medien
    Digitale Medien
    Springer
    Journal of inclusion phenomena and macrocyclic chemistry 10 (1991), S. 305-312 
    ISSN: 1573-1111
    Schlagwort(e): Crystallization ; adductive crystallization ; nucleation ; crystal growth ; polymorph ; transformation ; release rate ; perfume
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Chemie und Pharmazie
    Notizen: Abstract The precipitation behavior of 1,1-di(p-hydroxyphenyl)cyclohexane (DHC) from acetone solutions containing d-Limonene (1-methyl-4(I-methylethenyl)cyclohexene) was studied. From the pure acetone solution or the solutions containing a small amount of d-Limonene crystals (B) precipitated, which clathrate only acetone with a guest/host (G/H) molar ratio of 1.0. However, when thed-Limonene concentration is increased to more than ca. 2 mol/L, crystals (A) precipitated which had a different habit from the B crystals. In the A crystalsd-Limonene is clathrated together with a large amount of acetone and the G/H value ofd-Limonene increases with the concentration in the solution up to the maximum value of 0.2. As the diffraction patterns of the A and B crystals are similar, it is assumed that a part of the acetone molecules in the B crystals are replaced byd-Limonene molecules. The acetone in the A crystals escapes rapidly, but thed-Limonene remains for a long time. This may indicate that the large molecule ofd-Limonene cannot diffuse rapidly within the host lattice owing to three-dimensional hindrance. It was clear that the solubility of the A crystals is higher than that of the B crystals and the transformation from the'metastable A to the stable B crystals proceeds during the crystallization of A crystals.
    Materialart: Digitale Medien
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  • 276
    Digitale Medien
    Digitale Medien
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 45 (1991), S. 22-29 
    ISSN: 0730-2312
    Schlagwort(e): transformation ; tumor suppressor genes ; oncogenic mutations ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Although the case for p53 as a tumor suppressor gene appears very strong, one should still keep an open eye for the possibility that mutations in p53 do not necessarily imply a mere loss of “suppressor” activity. It is still possible that the presence of a p53 mutation in a tumor contributes, in a dominant positive manner, to tumorigenesis. In other words, certain p53 mutants may well be oncogenic in their own right, and carry distinct activities that promote growth deregulation and malignant progression. Elucidating this issue also has practical implications, since the nature of the resident mutations may greatly dictate the consequences of attempts to reintroduce wild-type (wt) p53 into particular types of tumor cells. There are two major obstacles along the road to meaningful answers: the limitations of the experimental systems used for evaluating the biological activities of Wt and mutant p53 and a fundamental lack of knowledge about the relevant biochemistry of the p53 protein. These two aspects constitute primary experimental challenges for investigators in the field.
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    Materialart: Digitale Medien
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  • 277
    Digitale Medien
    Digitale Medien
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 46 (1991), S. 277-283 
    ISSN: 0730-2312
    Schlagwort(e): transformation ; malignancy ; metastasis ; gene regulation ; cancer ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Changes in the quantitative expression of certain genes or in the amounts of their products can quickly stimulate progression to the metastatic phenotype. This has been done experimentally by transferring dominantly acting oncogenes such as c-H-rasEJ into susceptible cells or more recently by interfering with metastasis suppressor genes. In vivo such rapid qualitative changes in dominantly acting oncogenes or suppressor genes occur only rarely, and progression to highly metastatic phenotypes is thought to occur through a process involving the slow stepwise progression of a subpopulation of neoplastic cells to more malignant states. Such slow changes can be reversible and need not involve known dominantly acting oncogenes or metastatic suppressor genes, consistent with clinical and experimental observations on naturally occuring, highly advanced metastatic tumors. An important element in the natural progression of tumors to more malignant states may be their ability to circumvent host environmental controls that regulate growth and cellular diversity. They also evolve into heterogeneous cellular phenotypes, a process that appears to mainly involve quantitative changes in gene expression but can be rapidly stimulated in cell culture by the introduction of a dominantly acting oncogene or inhibited by the introduction of a suppressor gene. The oncogenes and suppressor genes that affect malignancy may control important steps in the quantitative regulation of sets of genes that are ultimately responsible for the cellular alterations seen in adhesion receptors, cell motility responses, cell-cell communication components, degradative enzymes and their inhibitors, growth factor receptors, components that aid in escape from host surveillance mechanisms and others that are important in malignancy. Highly malignant cells that have slowly evolved in vivo may contain only a few qualitative gene changes but have undergone extensive cycles of diversification and accumulation of quantitative changes in the expression of genes that encode products that are related to malignancy and metastasis. Thus highly malignant cells can arise quickly due to specific qualitative changes in critical controlling genes or more slowly by less critical qualitative genetic changes together with cycles of cellular diversification and accumulation of quantitative changes in gene expression.
    Zusätzliches Material: 1 Ill.
    Materialart: Digitale Medien
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  • 278
    Digitale Medien
    Digitale Medien
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 46 (1991), S. 260-265 
    ISSN: 0730-2312
    Schlagwort(e): rough membranes ; smooth membranes ; structural transitions ; transformation ; spin probes ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: This paper deals with microviscosity parameters and thermoinduced structural transitions in the lipids of smooth and heavy rough endoplasmic reticulum membranes isolated from Krebs II ascites cells incubated with the phorbol ester 12-0-tetradecanoyl-phorbol-13-acetate. The phorbol ester was found to bring about a threefold increase in the microviscosity of the lipids in heavy rough membranes. Spin probe I (2,2,6,6-tetrahydro-4-capryloyl-oxypiperidine-1-oxyl), localized in the surface layer of the membrane lipids, gave results which indicate an increased number of thermoinduced structural transitions in the smooth membranes in the treated cells due to the transitions occurring at relatively low temperature and a decreased number of such transitions in the heavy rough fraction especially at high temperature. For 5,6-benzo-2,2,4,4-tetramethyl-1,2,3,4-tetrahydro-γ-carboline-oxyl, probe II, mainly distributed in the annular lipids, a decrease in the number of low temperature transitions in the smooth fraction was observed, while an increase occurred in the heavy rough one. The results obtained are discussed in terms of the effect of phorbol esters as promoters of tumor progression.
    Zusätzliches Material: 3 Ill.
    Materialart: Digitale Medien
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  • 279
    ISSN: 0192-253X
    Schlagwort(e): cAMP ; chemotaxis ; transformation ; CAT constructs ; gene regulation ; Life and Medical Sciences ; Genetics
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: The genes coding for the cyclic nucleotide phosphodiesterase (PD) and the PD inhibitory glycoprotein (PDI) have been cloned and characterized. The PDI gene was isolated as a 1.6 kb genomic fragment, which included the coding sequence containing two small introns and 510 nucleotides of non-translated 5′ sequence. From the deduced amino acid sequence we predict a protein with a molecular weight (MW) of 26,000 that, in agreement with previous data, contains 15% cysteine residues. Genomic Southern blot analysis indicates that only one gene encodes the inhibitor. Northern blot analysis shows a single transcript of 0.95 kb. The PDI gene is expressed early in development with little transcript remaining following aggregation. The appearance of PDI mRNA is prevented by the presence of cAMP, but when cAMP is removed the transcript appears within 30 minutes. When cAMP is applied to cells expressing PDI the transcript disappears with a half-life of less than 30 minutes. The PD gene of D. discoideum is transcribed into three mRNAs: a 1.9 kb mRNA specific for growth, a 2.4 kb mRNA specific for aggregation, and a 2.2 kb mRNA specific for late development. The 2.2 kb mRNA is also specific for prestalk cells, and is induced by differentiation-inducing factor. All three mRNAs contain the same coding sequence, and differ only in their 5′ non-coding sequences. Each mRNA is transcribed from a different promoter, and by using the chloramphenicol acyltransferase gene as a reporter, we have shown that each promoter displays the same regulation as its cognate mRNA. Transformation of wild-type strains with the PD gene causes PD overexpression which accelerates aggregation and blocks subsequent cell differentiation and pattern formation.
    Zusätzliches Material: 8 Ill.
    Materialart: Digitale Medien
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  • 280
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell reports 9 (1990), S. 26-29 
    ISSN: 1432-203X
    Schlagwort(e): Solanum melongena ; eggplant ; Agrobacterium tumefaciens ; kanamycin ; transformation ; shoot regeneration
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Kanamycin resistant plants of Solarium melongena L. (eggplant) cv. Picentia were obtained following the cocultivation of leaf explants with Agrobacterium tumefaciens. A disarmed binary vector system containing the neomycin phosphotransferase (NPTII) gene as the selectable marker and chloramphenicol acetyltransferase (CAT) as a reporter gene was utilized. In vitro grown plants were used as sources of explants to produce transgenic plants on selective medium containing 100 mg/l kanamycin. The transformation and expression of the foreign genes was confirmed by DNA hybridizations, leaf disc assays, and by measuring NPTII and CAT enzyme activities. This technique is simple, rapid, efficient, and transgenic eggplants of this commercial cultivar have been transferred to soil where they have flowered and set seed.
    Materialart: Digitale Medien
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  • 281
    ISSN: 1573-5028
    Schlagwort(e): direct gene transfer ; gene rearrangements ; Nicotiana tabacum ; particle gun ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Transgenic tobacco plants and progeny carrying coding sequences for neomycin phosphotransferase II (NPTII) and beta-glucuronidase (GUS) were recovered following microprojectile bombardment of tobacco leaves. Transgenic plants were regenerated from bombarded leaf pieces of tobacco cvs. ‘Xanthi’ and ‘Ky 17’ which were cultured in the presence of 100 or 200 μg/ml kanamycin for six to eight weeks. Among 160 putative transgenic plants from at least 16 independent transformation events 76% expressed NPTII, and 50% expressed GUS. Southern analysis of plants expressing either one or both of the enzymes indicated DNA in high molecular weight DNA in 8 of 9 independent transformants analyzed. Two independent transformants and their progeny were analyzed in detail. Analysis of progeny for quantitative enzyme levels of NPTII and GUS, and Southern analysis of parents and progeny clearly demonstrated that the genes were transmitted to progeny. One transformant demonstrated Mendelian ratios for seed germination on kanamycin-containing medium while the other transformant had non-Mendelian ratios. DNA analysis of progeny indicate complex integration of the plasmid DNA, and suggest that rearrangements of this DNA has occurred. These results are consistent with other methods of direct DNA uptake into cells, and verify that the microprojectile bombardment method is capable of DNA delivery into intact plant cells which can give rise to transgenic plants and progeny.
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  • 282
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium ; conifers ; DNA hybridization ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The hypervirulent Agrobacterium tumefaciens strain A281 formed frequent tumors (31%) on Picea abies (Norway spruce), an economically important tree species in Swedish forests. Three-month-old seedlings were inoculated and tumors were established that grew hormone-independently in culture. Tumors contained agropine and mannopine/mannopinic acid as determined by acid pH paper electrophoresis. In addition, DNA hybridization studies showed that the DNA from these tumor lines contained sequences homologous to Ti plasmid T-DNA, whereas wild-type spruce seedling DNA did not. These results suggest that Agrobacterium vectors can be used for gene transfer into this important forest species.
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  • 283
    ISSN: 1573-5028
    Schlagwort(e): cytokinin ; tumor formation ; isopentenyltransferase gene ; Nicotiana ; genetic tumors ; polymerase chain reaction (PCR) ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The shooty morphology of a nontumorous amphidiploid mutant of Nicotiana glauca Grah. x N. langsdorffii Weinm. was restored by cytokinins, whether exogenously applied or endogenously produced by transformation of the mutant with a transfer DNA (T-DNA) cytokinin-biosynthesis gene (isopentenyltransferase; ipt). Auxins alone did not confer this effect. Similar transformation was not achieved for the parental species. In the case of transformation with the ipt gene, selection of the transformed tissues was based on its hormone-independent growth in the presence of the antibiotic kanamycin. Transformed tissues exhibited a shooty morphology, indistinguishable from that of wildtype genetic tumors N. glauca x N. langsdorffii. This altered phenotype was caused by the presence and constitutive expression of the ipt gene. The insertion and expression of this gene in transformed tissues was confirmed by using the polymerase chain reaction (PCR) technique as well as conventional molecular hybridization analysis. Expression of the ipt gene led to an elevated level of cytokinin in the transformed mutant tissues. This evidence supports the notion that genetic tumors are caused, at least in part, by elevated levels of cytokinin in interspecific hybrids.
    Materialart: Digitale Medien
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  • 284
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 14 (1990), S. 785-792 
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium rhizogenes ; Agrobacterium tumefaciens ; root induction ; tzs ; Linum usitatissimum L. ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Root induction on flax (Linum usitatissimum L.) cotyledon explants by Agrobacterium rhizogenes strain 1855 is markedly increased by co-inoculation with disarmed A. tumefaciens strain LBA 4404 containing a plasmid carrying the tzs gene of pTiC58. Most of the roots (estimated to be more than 90%) were transformed. This effect is most likely due to the secretion of trans-zeatin by A. tumefaciens stimulating the division of plant cells making them more receptive to transformation by A. rhizogenes, although other explanations are possible. This observation supports the idea that the tzs gene, although not essential for transformation, may promote transformation. An obvious application for genetic engineering experiments involving transformation by A. rhizogenes, is to include a vir-induced tzs gene in the transformation system to help maximize transformation efficiency.
    Materialart: Digitale Medien
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  • 285
    ISSN: 1573-5028
    Schlagwort(e): antisense ; gene expression ; plants ; regulation ; RNA ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have shown leaf-specific inhibition GUS gene expression in transgenic Nicotiana plants using an antisense RNA with a 41-base homology spanning the translation start codon of the gene. GUS was expressed from the nominally constitutive 35S promoter and the antisense RNA was expressed from the light-regulated ca/b promoter of Arabidopsis thaliana. A range of GUS inhibition from 0 to 100% was obtained by screening a small population of transgenic plants and the specific levels of inhibition observed were stably inherited in two generations. An antiGUS ‘gene’ dosage effect was observed in plants which were homozygous for antiGUS. RNA detection results suggest that duplex formation with the 41 base pair antiGUS RNA destabilized the GUS mRNA and that an excess of antisense. RNA was not required. Our results demonstrate the potential of antisense RNA as a strategy for obtaining plant mutants, especially ‘down mutations’ in essential genes where only a short 5′ sequence of the mRNA is required. They also suggest that the ‘position effect’ on gene expression could be used in conjunction with an antisense RNA strategy to provide a versatile approach for crop improvement.
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  • 286
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 14 (1990), S. 815-824 
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium tumefaciens ; hypocotyls ; Nicotiana alata ; regeneration ; self-incompatibility ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A transformation and regeneration system has been developed for Nicotiana alata, a plant which is being intensively studied as a model of gametophytic self-incompatibility. Plantlets can be regenerated efficiently from seedling hypocotyls. Kanamycin-resistant, transformed plants have been obtained by cocultivation of regenerating hypocotyls with Agrobacterium tumefaciens strain LBA4404 containing a binary vector. The transformation frequency was low with 〈1% of tissue explants regenerating transformed plants. The transformed plants contained from one to three copies of the introduced DNA. In most cases, the kanamycin resistance phenotype was transmitted to the offspring as a normal Mendelian factor. In one unusual case, none of the offspring inherited the kanamycin resistance of the transformed maternal parent. This plant may have been chimeric or the kanamycin resistance gene may have been inactivated.
    Materialart: Digitale Medien
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  • 287
    Digitale Medien
    Digitale Medien
    Springer
    Cancer and metastasis reviews 9 (1990), S. 93-98 
    ISSN: 1573-7233
    Schlagwort(e): transformation ; progression ; autotrophs
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract In 1984, Philippe Shubik wrote in an editorial in the Journal of the National Cancer Institute [1] that: ‘In terms of general biology, the multicellular organisms have evolved a complex series of defensive responses to extracellular injury involving various inflammatory reactions and their systemic concomitants. Needless to say, these reactions are by no means always successful from the standpoint of the host and indeed may well be the immediate cause of the obvious ill effects noted. The unicellular organisms, in contrast, react to injury by dividing and moving. In the search for the features that may link chemical, physical, and viral carcinogens apart from their ability to induce neoplasia, only one characteristic in common is obvious, namely, their ability to produce intracellular change or injury while leaving the cell viable. Perhaps the initial and fundamental characteristic of neoplasia is a reversion of the cell to unicellular behavior. Division and invasiveness are the characteristics of the neoplastic cell, and increased motility certainly seems to be the most likely mechanism for invasion’. Dr. Shubik presented these views as a basis for ‘further discussion’ regarding the nature of the neoplastic response. We hope that this presentation will augment Shubik's plea by reviewing his idea in the context of our current knowledge of tumor development. In addition, we will attempt to integrate the concept of the unicellular behavior of tumor cells with Foulds' [2] and subsequently Nowell's [3] insightful hypothesis concerning tumor progression.
    Materialart: Digitale Medien
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  • 288
    Digitale Medien
    Digitale Medien
    Springer
    Plant foods for human nutrition 40 (1990), S. 283-288 
    ISSN: 1573-9104
    Schlagwort(e): heat ; transformation ; fatty acids ; leaf proteins
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft
    Notizen: Abstract Freeze-dried leaf protein concentrate (LPC) contained 18% lipids in which linolenic acid (61.5%) was the major component. Linolenic acid in LPC was almost stable when stored at ambient temperature (30 to 35°C) and exposed to air for 24 weeks. Heating of LPC (50 to 200°C) in presence of moisture (6 to 12%) progressively increased the rate of destruction of linolenic acid. Below 100°C the presence of lipids did not affect the protein quality but at higher temperatures due to the lipid oxidation protein quality as estimated by dye-binding capacity was considerably affected.
    Materialart: Digitale Medien
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  • 289
    ISSN: 1572-994X
    Schlagwort(e): HPV 16 ; transformation ; NIH 3T3 cells ; polycistronic mRNA ; cDNA cloning ; sequencing
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract We have cloned cDNA of the major 1.8 kb mRNA from HPV 16-transformed NIH 3T3 cells (PM3T3). The entire nucleotide sequences of this cDNA were determined and compared with prototype HPV 16 genomic DNA sequences. The 5′-end of the cDNA was flanked by approximately 300 bp of cellular sequences, and the 3′-end of the cDNA sequences contained poly A residues following at nt 4230. HPV 16 sequences began at nt 124, downstream of a major viral p97 promoter, within the E6 open reading frame (ORF). The first splice donor site was at nt 226 and the splice acceptor site was at nt 409, suggesting that the E6 gene is inert. Second splice donor and acceptor sites were located at nt 880 and at nt 3357, respectively. This mRNA was thus shown to consist of three exons, resulting in polycistronic mRNA containing three potentially functional virus early genes-E7, E1^E4, and E5-actively transcribed in the transformant.
    Materialart: Digitale Medien
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  • 290
    Digitale Medien
    Digitale Medien
    Springer
    Plant cell, tissue and organ culture 22 (1990), S. 135-145 
    ISSN: 1573-5044
    Schlagwort(e): co-transformation ; liposomes ; tobacco ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract A system for the transformation of tobacco mesophyll protoplasts using pH-sensitive liposomes was developed. Plasmid DNA (plGVneo23) encoding the NPT-II gene for kanamycin resistance was entrapped in pH-sensitive liposomes composed of dioleolphosphatidylethanolamine, cholesterol and oleic acid. These liposomes release their contents at low pH and are capable of delivering their contents into the cytoplasm of protoplasts. Kanamycin-resistant colonies were reproducibly recovered from transformed protoplasts at an average frequency of 1.62×10-4 at pH 7.5. Plants regenerated from transformed cell lines were normal in appearance and were fertile. NPT-II activity was detected in leaf extracts of transformed, kanamycin-resistant plants and the presence of NPT-II DNA in the tobacco genome was shown by Southern blots. Analysis of self-pollinations and reciprocal crosses to non-transformed plants indicated that kanamycin resistance segregated as a dominant nuclear marker. Co-transformation of protoplasts with liposomes containing two selectable markers indicated that co-transformation occurred with a frequency of approximately 23%.
    Materialart: Digitale Medien
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  • 291
    Digitale Medien
    Digitale Medien
    Springer
    Cell biology and toxicology 6 (1990), S. 353-363 
    ISSN: 1573-6822
    Schlagwort(e): Transforming growth factor β1 ; cell cycle ; liver ; epithelial cell line ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: The effects of TGFβ1 on cell cycle events in a rat liver derived epithelial cell line (BL9) and in two in vitro transformants of this line were studied by flow cytometry. Using either ethidium bromide staining or the incorporation of bromodeoxyuridine to evaluate DNA synthesis it was shown that TGFβ1 prevented the entry of G0/G1 phase BL9 cells into S phase. TGFβ1 did not exert its inhibitory effect(s) on DNA synthesis by the modulation of early events in the cell cycle. The tumorigenic transformed BL9 cell lines gave contrasting responses to the effects of TGFβ1. DNA synthesis in a BL9 cell line derived by transfection with an active N-ras oncogene was unaffected by TFGβ1 and thus appeared refractory to its growth controlling effects. On the other hand cells from a BL9 cell line derived by in vitro transformation with activated aflatoxin B1 retained their sensitivity to the effects of TGFβ1. Thus the loss of the inhibitory effect of TGFβ1 on DNA synthesis is not obligatory for the malignant transformation of rat liver epithelial cells.
    Materialart: Digitale Medien
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  • 292
    Digitale Medien
    Digitale Medien
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 391-395 
    ISSN: 0192-253X
    Schlagwort(e): Lipofectin ; slime mould ; transformation ; Life and Medical Sciences ; Genetics
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie
    Notizen: We have studied the transient expression, in Dictyostelium cells growing on a bacterial food source, of a construct containing the coding region of the firefly luciferase gene inserted downstream of a Dictyosteliumactin promoter. The fusion gene is not detectably expressed when DNA is introduced by calcium phosphate precipitation or by electroporation, but it is expressed when introduced using cationic liposomes (lipofectin). Using this latter procedure, we are able to transform cells with a G418 resistance vector and select stable, drug-resistant transformants at a relatively low, but workable, efficiency. This technique will allow molecular genetics to be applied to the many important nonaxenic Dictyostelium strains.
    Zusätzliches Material: 2 Ill.
    Materialart: Digitale Medien
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  • 293
    Digitale Medien
    Digitale Medien
    Springer
    Current genetics 16 (1989), S. 369-372 
    ISSN: 1432-0983
    Schlagwort(e): Bialaphos ; Neurospora crassa ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
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  • 294
    ISSN: 1436-6215
    Schlagwort(e): γ-Tocopherol ; Transformation ; α-Tocopherol ; Ratten ; Generationsversuch ; γ-tocopherol ; transformation ; α-tocopherol ; rats ; generation-experiment
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Land- und Forstwirtschaft, Gartenbau, Fischereiwirtschaft, Hauswirtschaft , Medizin
    Beschreibung / Inhaltsverzeichnis: 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.
    Notizen: 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.
    Materialart: Digitale Medien
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  • 295
    Digitale Medien
    Digitale Medien
    Springer
    Methods in cell science 12 (1989), S. 127-133 
    ISSN: 1573-0603
    Schlagwort(e): polyethylene glycol ; transformation ; protoplasts ; Nicotiana
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
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  • 296
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium rhizogenes ; border sequence ; T-DNA ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
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  • 297
    ISSN: 1573-4943
    Schlagwort(e): conformational energy ; amino acid substitution ; P21 protein ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Chemie und Pharmazie
    Notizen: 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.
    Materialart: Digitale Medien
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  • 298
    Digitale Medien
    Digitale Medien
    Springer
    Plant molecular biology 13 (1989), S. 151-161 
    ISSN: 1573-5028
    Schlagwort(e): β-glucuronidase (gusA) gene ; maize ; protoplasts ; stable co-transformation ; transformation ; Zea mays L.
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
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  • 299
    ISSN: 1573-5028
    Schlagwort(e): Agrobacterium tumefaciens ; adventitious shoot regeneration ; transformation ; homozygous potato
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
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  • 300
    ISSN: 1573-5028
    Schlagwort(e): herbicide ; mutant ; photosystem II ; psbA gene ; transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
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