Abstract
Protoplasts prepared from cultured tobacco cells were treated with ColE1-kan plasmid DNA, a hybrid of ColE1 and pSC105 plasmids bearing a gene for kanamycin resistance. The conditions employed permitted the uptake or irreversible binding of 2.9% of the added DNA in acid-insoluble form. Upon commencement of division, the treated cells were plated in agar medium containing kanamycin and differentiating hormones. Plantlets or shoots obtained as presumptive transformants were further tested on kanamycin medium by subculturing small leaf pieces. No evidence was obtained for expression of the kanamycin resistance gene of ColE1-kan in tobacco tissue.
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Selected References
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- Aoki S., Takebe I. Infection of tobacco mesophyll protoplasts by tobacco mosaic virus ribonucleic acid. Virology. 1969 Nov;39(3):439–448. doi: 10.1016/0042-6822(69)90092-0. [DOI] [PubMed] [Google Scholar]
- Cocking E. C. Uptake of foreign genetic material by plant protoplasts. Int Rev Cytol. 1977;48:323–343. doi: 10.1016/s0074-7696(08)61748-9. [DOI] [PubMed] [Google Scholar]
- Hershfield V., Boyer H. W., Yanofsky C., Lovett M. A., Helinski D. R. Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455–3459. doi: 10.1073/pnas.71.9.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hinnen A., Hicks J. B., Fink G. R. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. doi: 10.1073/pnas.75.4.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hughes B. G., White F. G., Smith M. A. Fate of bacterial plasmid DNA during uptake by barley protoplasts. FEBS Lett. 1977 Jul 1;79(1):80–84. doi: 10.1016/0014-5793(77)80355-4. [DOI] [PubMed] [Google Scholar]
- Lurquin P. F., Kado C. I. Escherichia coli plasmid pBR313 insertion into plant protoplasts and into their nuclei. Mol Gen Genet. 1977 Jul 20;154(2):113–121. doi: 10.1007/BF00330826. [DOI] [PubMed] [Google Scholar]
- Uchimiya H., Murashige T. Evaluation of parameters in the isolation of viable protoplasts from cultured tobacco cells. Plant Physiol. 1974 Dec;54(6):936–944. doi: 10.1104/pp.54.6.936. [DOI] [PMC free article] [PubMed] [Google Scholar]