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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 9 (1990), S. 168-172 
    ISSN: 1432-203X
    Keywords: Oryza sativa L. ; indica rice, cotransformation ; protoplast ; kanamycin ; β ; glucuronidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts of the indica rice (Oryza sativa L.) variety, IR54, were transiently transformed with the gusA gene and stably transformed with both the neo and gusA genes. We show that PEG-mediated co-transformation of protoplasts with two genes on separate plasmids coupled with selection on kanamycin is an effective way of transferring foreign gene(s) into the indica rice genome. The efficiency of co-transformation was generally 20–30%, i.e. the frequency of kanamycin-resistant calli having both the neo and gusA active genes. Southern blot analysis using a probe for gusA indicated integration of several copies of the gene, often as head to tail tandem repeats.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 83 (1992), S. 855-863 
    ISSN: 1432-2242
    Keywords: Transformation ; PEG ; Indica rice ; Protoplast culture and regeneration ; neo ; gusA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have established a system to genetically engineer indica rice plants. In order to obtain transgenic plants, genes were introduced into protoplasts isolated from suspension cells of the indica rice var ‘IR54’ with the aid of polyethylene glycol (PEG). The neo gene was on pKAN and the gusA gene was on pPUR. The promoter for both genes was CaMV35S. Transformed calli were readily recovered from medium supplemented with G-418. In contrast, kanamycin interfered with plant regeneration from protoplast-callus. Transgenic plants were regenerated from calli resistant to G-418 in several separate experiments and grown to maturity in a growth chamber. Southern blot analysis of DNA isolated from leaves of T0 plants verified the presence of the transferred neo and gusA genes in the plant genome. A study of gene expression showed that the CaMV35SgusA gene was active in all of the organs examined. Mendelian inheritance of the introduced gusA gene was observed in progeny obtained by backcrossing the T0 plants to untransformed plants.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 27 (1995), S. 91-104 
    ISSN: 1573-5028
    Keywords: Oryza sativa L ; transgenic rice ; transgene inheritance ; gusA gene ; neo gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Inheritance of foreign genes neo and gusA in rice (Oryza sativa L. cv. IR54 and Radon) has been investigated in three different primary (T0) transformants and their progeny plants. T0 plants were obtained by co-transforming protoplasts from two different rice suspension cultures with the neomycin phosphotransferase II gene [neo or aph (3′) II] and the β-glucuronidase gene (uidA or gusA) residing on separate chimeric plasmid constructs. The suspension cultures were derived from callus of immature embryos of indica variety IR54 and japonica variety Radon. One transgenic line of Radon (AR2) contained neo driven by the CaMV 35S promoter and gusA driven by the rice actin promoter. A second Radon line (R3) contained neo driven by the CaMV 35S promoter and gusA driven by a promoter of the rice tungro bacilliform virus. The third transgenic line, IR54-1, contained neo driven by the CaMV 35S promoter and gusA driven by the CaMV 35S. Inheritance of the transgenes in progeny of the transgenic rice was investigated by Southern blot analysis and enzyme assays. Southern blot analysis of genomic DNA showed that, regardless of copy numbers of the transgenes in the plant genome and the fact that the two transgenes resided on two different plasmids before transformation, the introduced gusA and neo genes were stably transmitted from one generation to another and co-inherited together in transgenic rice progeny plants derived from self-pollination. Analysis of GUS and NPT II activities in T1 to T2 plants provided evidence that inheritance of the gusA and neo genes was in a Mendelian fashion in one plant line (AR2), and in an irregular fashion in the two other plant lines (R3 and IR54-1). Homozygous progeny plants expressing the gusA and neo genes were obtained in the T2 generation of AR2, but the homozygous state was not found in the other two lines of transgenic rice.
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  • 4
    Publication Date: 2021-10-01
    Print ISSN: 0360-3199
    Electronic ISSN: 1879-3487
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Published by Elsevier
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