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  • 1985-1989  (2)
  • MgCl2-PEG-electroporation  (1)
  • Nicotiana plumbaginifolia  (1)
  • 1
    Electronic Resource
    Electronic Resource
    Effect of repeated DNA sequences on direct gene transfer in protoplasts of Nicotiana plumbaginifolia (1989)
    Springer
    Theoretical and applied genetics 78 (1989), S. 113-118 
    Details
    ISSN: 1432-2242
    Keywords: Nicotiana plumbaginifolia ; Protoplasts ; Gene transfer ; Repeated DNA sequences
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Highly repeated nuclear DNA sequences from leaves of Nicotiana plumbaginifolia were cloned in pBR322 and tested for their effect on direct gene transfer in protoplasts of the same organism. Protoplasts were prepared from suspension cultures and were incubated in the presence of the plasmid pHP23 carrying the kanamycin resistance gene APH(3′)II and in the presence of the plasmids carrying the cloned sequence. DNA uptake was induced by a polyethyleneglycol (PEG) treatment. Out of the 22 tested clones, 3 significantly stimulated the frequency of appearance of transformed colonies. DNA was extracted from some of the kanamycin-resistant calli obtained by co-transformations. Dot-blots have shown that the stimulatory effect on transformation frequency is often accompanied by a consistent increase in integrated genes sequences.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00299763
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Hybrid genes in the analysis of transformation conditions (1987)
    Springer
    Plant molecular biology 8 (1987), S. 363-373 
    Details
    ISSN: 1573-5028
    Keywords: transformation ; MgCl2-PEG-electroporation ; competence ; protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Direct gene transfer into plant protoplasts has been recently developed, and conditions for high frequency transformation of SR1 tobacco protoplasts established. In this paper we analyse numerous transformation parameters in a comparative study on SR1 Nicotiana tabacum and N. plumbaginifolia, and report on a simple chemical technique for very efficient protoplast transformation. It is based on the synergistic interaction of MgCl2 and PEG. The technique yielded up to 1400 transformants per 3×105 treated N. tabacum protoplasts (up to 4.8% of the survivors, late selected clones). Using N. plumbaginifolia, the frequencies were 10-fold lower, indicating that the ‘competence’ for transformation has a species-specific component.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00015814
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    Paper (German National Licenses)
    Fulltext
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