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  • 1
    Electronic Resource
    Electronic Resource
    Transgenic Medicinal Plants: Agrobacterium-Mediated Foreign Gene Transfer and Production of Secondary Metabolites (1992)
    s.l. : American Chemical Society
    Journal of natural products 55 (1992), S. 149-162 
    Details
    ISSN: 1520-6025
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1021/np50080a001
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  • 2
    Electronic Resource
    Electronic Resource
    Transgenic fertile Scoparia dulcis L., a folk medicinal plant, conferred with a herbicide-resistant trait using an Ri binary vector (1996)
    Springer
    Plant cell reports 15 (1996), S. 317-321 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transgenic herbicide-resistant Scoparia dulcis plants were obtained by using an Ri binary vector system. The chimeric bar gene encoding phosphinothricin acetyltransferase flanked by the promoter for cauliflower mosaic virus 35S RNA and the terminal sequence for nopaline synthase was introduced in the plant genome by Agrobacterium-mediated transformation by means of scratching young plants. Hairy roots resistant to bialaphos were selected and plantlets (R0) were regenerated. Progenies (S1) were obtained by self-fertilization. The transgenic state was confirmed by DNA-blot hybridization and assaying of neomycin phosphotransferase II. Expression of the bar gene in the transgenic R0 and S1 progenies was indicated by the activity of phophinothricin acetyltransferase. Transgenic plants accumulated scopadulcic acid B, a specific secondary metabolite of S. dulcis, in amounts of 15–60% compared with that in normal plants. The transgenic plants and progenies showed resistant trait towards bialaphos and phosphinothricin. These results suggest than an Ri binary system is one of the useful tools for the transformation of medicinal plants for which a regeneration protocol has not been established.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050024
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  • 3
    Electronic Resource
    Electronic Resource
    Transgenic herbicide-resistant Atropa belladonna using an Ri binary vector and inheritance of the transgenic trait (1992)
    Springer
    Plant cell reports 11 (1992), S. 219-224 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transgenic Atropa belladonna conferred with a herbicide-resistant trait was obtained by transformation with an Ri plasmid binary vector and plant regeneration from hairy roots. We made a chimeric construct, pARK5, containing the bar gene encoding phosphinothricin acetyltransferase flanked with the promoter for cauliflower mosaic virus 35S RNA and the 3′ end of the nos gene. Leaf discs of A. belladonna were infected with Agrobacterium rhizogenes harboring an Ri plasmid, pRi15834, and pARK5. Transformed hairy roots resistant to bialaphos (5 mg/l) were selected and plantlets were regenerated. The integration of T-DNAs from pRi15834 and pARK5 were confirmed by DNA-blot hybridization. Expression of the bar gene in transformed R0 tissues and in backcrossed F1 progeny with a nontransformant and self-fertilized progeny was indicated by enzymatic activity of the acetyltransferase. The transgenic plants showed resistance towards bialaphos and phosphinothricin. Tropane alkaloids of normal amounts were produced in the transformed regenerants. These results present a successful application of transformation with an Ri plasmid binary vector for conferring an agronomically useful trait to medicinal plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00235069
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  • 4
    Electronic Resource
    Electronic Resource
    Transgenic fertile Scoparia dulcis L., a folk medicinal plant, conferred with a herbicide-resistant trait using an Ri binary vector (1996)
    Springer
    Plant cell reports 15 (1996), S. 317-321 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transgenic herbicide-resistant Scoparia dulcis plants were obtained by using an Ri binary vector system. The chimeric bar gene encoding phosphinothricin acetyltransferase flanked by the promoter for cauliflower mosaic virus 35S RNA and the terminal sequence for nopaline synthase was introduced in the plant genome by Agrobacterium-mediated transformation by means of scratching young plants. Hairy roots resistant to bialaphos were selected and plantlets (R0) were regenerated. Progenies (S1) were obtained by self-fertilization. The transgenic state was confirmed by DNA-blot hybridization and assaying of neomycin phosphotransferase II. Expression of the bar gene in the transgenic R0 and S1 progenies was indicated by the activity of phosphinothricin acetyltransferase. Transgenic plants accumulated scopadulcic acid B, a specific secondary metabolite of S. dulcis, in amounts of 15–60% compared with that in normal plants. The transgenic plants and progenies showed resistant trait towards bialaphos and phosphinothricin. These results suggest that an Ri binary system is one of the useful tools for the transformation of medicinal plants for which a regeneration protocol has not been established.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232363
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  • 5
    Electronic Resource
    Electronic Resource
    Tissue-specific and stress-enhancing expression of the TR promoter for mannopine synthase in transgenic medicinal plants (1991)
    Springer
    Planta 184 (1991), S. 40-46 
    Details
    ISSN: 1432-2048
    Keywords: Digitalis ; Gene expression ; Glycyrrhiza ; Nicotiana (transgenic) ; TR promoter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic tobacco (Nicotiana tabacum L.), licorice (Glycyrrhiza uralensis Fisher) and foxglove (Digitalis purpurea L.) were obtained with binary vector systems based on a disarmed Agrobacterium tumefaciens and on a virulent A. rhizogenes. The chimeric neomycin phosphotransferase II (NPT-II) gene (kan) and the ß-glucuronidase (GUS) gene (uidA) were under the control of the TR1′ and 2′ promoters, respectively, on a binary vector, pGSGluc1. Tissue-specific expression of the chimeric TR2' -uidA gene was studied using regenerants of N. tabacum from transformed callus and hairy roots, and also using the transformed roots of G. uralensis and D. purpurea. In all these transformed tissues, the phloem and surrounding tissues were stained with 5-bromo-4-chloro-3-indolyl-ß-d-glucuronide, showing the specific expression of uidA. The enzymatic assays of NPT-II and GUS indicated that the expression of both TR1' -kan and TR2'- uidA were coordinately enhanced by wounding and by the addition of plant growth regulators. These results indicate that the gene expression by the dual TR promoters is regulated in the several plant species studied in a tissue-specific manner and enhanced by physiological stresses.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00208234
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  • 6
    Electronic Resource
    Electronic Resource
    Genetic transformation of foxglove (Digitalis purpurea) by chimeric foreign genes and production of cardioactive glycosides (1990)
    Springer
    Plant cell reports 9 (1990), S. 121-124 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The chimeric neo and gus genes on a mini Ti vector are efficiently transferred into the genome of fox glove (Digitalis purpurea L.) using a binary vector system based on a rootinducing Ri plasmid, pRi15834. The transgenic state of established transformed roots was confirmed by Southern blot analysis and by detection of agropine and mannopine. The expression of the chimeric genes controlled by the promoters from TR 1′–2′ genes, nos gene and cauliflower mosaic virus 35S RNA was demonstrated by enzymatic and histochemical assays of neomycin phosphotransferase II and ß-glucuronidase. Enzyme-linked immunosorbent assay (ELISA) was carried out using polyclonal antibody reactable against digitoxin to investigate the production of cardenolides. The results of ELISA indicated that the cardioactive glycosides were highly produced in the green transformed hairy roots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232085
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  • 7
    Electronic Resource
    Electronic Resource
    Stable transfer and expression of chimeric genes in licorice (Glycyrrhiza uralensis) using an Ri plasmid binary vector (1990)
    Springer
    Plant cell reports 8 (1990), S. 718-721 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The pharmaceutically important plant, licorice (Glycyrrhiza uralenesis Fisher), was transformed with a binary vector system of an Ri plasmid, pRi15834, and a mini Ti vector, pGSGluc1, containing chimeric neo and gus genes. The transgenic state of transformed roots was confirmed by detection of agropine and mannopine and by Southern blot hybridization with T-DNA of pGSGluc1. One to three copies of T-DNA of pGSGluc1 was integrated into the genomic DNA of G. uralensis. The expression of chimeric neo and gus genes driven by TR 1′ and 2′ promoters, respectively, was demonstrated by enzymatic assays. Histochemical analysis showed that the chimeric TR2′-gus gene was expressed specifically in phloem and pericycle tissues of the transformed licorice roots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00272102
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  • 8
    Electronic Resource
    Electronic Resource
    A constitutively expressed Myc-like gene involved in anthocyanin biosynthesis from Perilla frutescens: molecular characterization, heterologous expression in transgenic plants and transactivation in yeast cells (1999)
    Springer
    Plant molecular biology 41 (1999), S. 33-44 
    Details
    ISSN: 1573-5028
    Keywords: anthocyanin biosynthesis ; basic helix-loop-helix protein ; Myc gene ; Perilla frutescens ; transactivation domain ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The coordinate expression of anthocyanin biosynthetic genes in leaves and stems of a red forma of Perilla frutescens is presumably controlled by regulatory gene(s). A Myc-like gene (Myc-rp) was isolated from a cDNA library prepared from the leaves of red P. frutescens, and its deduced amino acid sequence shows 64% identity with that of delila from snapdragon. The Myc-rp gene was expressed in leaves and roots of both red and green P. frutescens equally. Comparison of deduced amino acid sequence of Myc-rp with that of Myc-gp, the second allele isolated from a green forma of P. frutescens, indicates that the 132nd amino acid, alanine, existing in MYC-RP was changed to serine in MYC-GP. The heterologous expression of these two alleles of Myc-like gene in tobacco and tomato resulted in an increase of the anthocyanin contents in flowers of tobacco and vegetative tissues and flowers of tomato. However, the flowers of transgenic tobacco expressing the fragment with a partial deletion (encoding 1–115 amino acids deleted) of Myc-gp gave no change in anthocyanin accumulation, but some morphological changes of the flower were observed. In yeast, the MYC-RP/GP and Delila protein exhibited transactivation activity on the GAL-1 promoter from yeast and the promoter of dihydroflavonol 4-reductase (DFR) gene from P. frutescens. A transactivation domain of MYC-RP/GP and Delila could be located in the region between the 193rd and the 420th amino acid of MYC-RP/GP proteins. Our data indicate that this Myc-like gene presumably functions in the regulation of anthocyanin biosynthesis similarly in different tissues of dicot plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006237529040
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  • 9
    Electronic Resource
    Electronic Resource
    Molecular cloning and expression of key enzymes for biosynthesis of cysteine and related secondary non-protein amino acids (1994)
    Springer
    Plant cell, tissue and organ culture 38 (1994), S. 153-158 
    Details
    ISSN: 1573-5044
    Keywords: O-acetylserine (thiol) lyase ; cDNA cloning ; cysteine synthase ; genetic complementation ; Spinacia oleracea ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cysteine synthase plays a key role in the sulfur assimilation pathway in plant cells. The cDNA clones encoding two isoforms of this enzyme were isolated from spinach by synthetic oligonucleotide probes. The modes of expression of these two genes differed in tissues of spinach. A heterologous expression system in Escherichia coli and transgenic tobacco was made. The application of heterologous expression to modify sulfur metabolism and to produce non-protein amino acids is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00033872
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  • 10
    Electronic Resource
    Electronic Resource
    Cloning and molecular analysis of structural genes involved in anthocyanin biosynthesis and expressed in a forma-specific manner in Perilla frutescens (1997)
    Springer
    Plant molecular biology 35 (1997), S. 915-927 
    Details
    ISSN: 1573-5028
    Keywords: anthocyanin ; cDNA cloning ; forma-specific expression ; genomic organization ; light induction ; Perilla frutescens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two cultivars of Perilla frutescens, red and green formas, are known to differ in anthocyanin accumulation in leaves and stems. cDNA clones encoding the enzymes involved in anthocyanin biosynthesis, chalcone synthase (CHS), flavanone 3-hydroylase (F3H), dihydroflavonol 4-reductase (DFR), and UDP glucose: flavonoid 3-O-glucosyltransferase (3GT), were isolated from cDNA libraries derived from the leaves of a red forma of P. frutescens by screening with partial fragments amplified by means of polymerase chain reaction (PCR) and heterologous cDNAs as probes. The deduced amino acid sequences of these four genes exhibited 40–90% identity with those reported for the corresponding gene from other unrelated species. Southern blot analysis for these genes and two other structural genes, the leucoanthocyanidin dioxygenase (LDOX, anthocyanidin synthase) and anthocyanin acyltransferase (AAT) genes, indicated that each gene comprises a small multi-gene family. More than three copies of the CHS gene are present, two copies of the other genes being present. The expression of five genes, the exception being the CHS gene, was detected only in red leaves of the red forma of P. frutescens, i.e. not in green leaves of the green forma plant. The CHS gene was expressed in both red and green leaves, but 10-fold more in red leaves than in green leaves. These results suggest that the expression of all structural genes examined is coordinately regulated in a forma-specific manner. Under weak-light conditions, the accumulation of both anthocyanin and mRNAs of biosynthetic enzymes was lower in leaves of the red forma. High-intensity white light coordinately induced the accumulation of transcripts of all six genes examined in the mature leaves of red P. frutescens.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005959203396
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