Abstract
Protoplasts of Saccharomyces cerevisiae were mixed with linear DNA plasmids, pGKl1 and pGKl2, isolated from a Kluyveromyces lactis killer strain and treated with polyethylene glycol. Out of 2,000 colonies regenerated on a nonselective medium, two killer transformants were obtained. The pGKl plasmids and the killer character were stably maintained in one (Pdh-1) of them. Another transformant, Pdl-1, was a weak killer, and the subclones consisted of a mixture of weak and nonkiller cells. The weak killers were characterized by the presence of pGKl1 in a decreased amount, and nonkillers were characterized by the absence of pGKl1. The occurrence of two new plasmids which migrated faster than pGKl1 in an agarose gel was observed in Pdl-1 and its subclones, whether weak or nonkillers. Staining with 4',6-diamidino-2-phenylindole revealed that the pGKl plasmids exist in the cytosol of transformant cells with numerous copy numbers.
Full text
PDF


Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cameron J. R., Philippsen P., Davis R. W. Analysis of chromosomal integration and deletions of yeast plasmids. Nucleic Acids Res. 1977;4(5):1429–1448. doi: 10.1093/nar/4.5.1429. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chouikh Y., Volovitch M., Yot P. A simple and fast electrophoretic method for elution of nucleic acids from gels. Mol Biol Rep. 1979 Dec 31;5(4):237–239. doi: 10.1007/BF00782896. [DOI] [PubMed] [Google Scholar]
- Clark-Walker G. D., Miklos G. L. Localization and quantification of circular DNA in yeast. Eur J Biochem. 1974 Jan 16;41(2):359–365. doi: 10.1111/j.1432-1033.1974.tb03278.x. [DOI] [PubMed] [Google Scholar]
- Gunge N., Sakaguchi K. Intergeneric transfer of deoxyribonucleic acid killer plasmids, pGKl1 and pGKl2, from Kluyveromyces lactis into Saccharomyces cerevisiae by cell fusion. J Bacteriol. 1981 Jul;147(1):155–160. doi: 10.1128/jb.147.1.155-160.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gunge N., Tamaru A., Ozawa F., Sakaguchi K. Isolation and characterization of linear deoxyribonucleic acid plasmids from Kluyveromyces lactis and the plasmid-associated killer character. J Bacteriol. 1981 Jan;145(1):382–390. doi: 10.1128/jb.145.1.382-390.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Niwa O., Sakaguchi K., Gunge N. Curing of the killer deoxyribonucleic acid plasmids of Kluyveromyces lactis. J Bacteriol. 1981 Dec;148(3):988–990. doi: 10.1128/jb.148.3.988-990.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Petes T. D. Molecular genetics of yeast. Annu Rev Biochem. 1980;49:845–876. doi: 10.1146/annurev.bi.49.070180.004213. [DOI] [PubMed] [Google Scholar]
- Somers J. M., Bevan E. A. The inheritance of the killer character in yeast. Genet Res. 1969 Feb;13(1):71–83. doi: 10.1017/s0016672300002743. [DOI] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Wickner R. B. The killer double-stranded RNA plasmids of yeast. Plasmid. 1979 Jul;2(3):303–322. doi: 10.1016/0147-619x(79)90015-5. [DOI] [PubMed] [Google Scholar]
- Williamson D. H., Fennell D. J. The use of fluorescent DNA-binding agent for detecting and separating yeast mitochondrial DNA. Methods Cell Biol. 1975;12:335–351. doi: 10.1016/s0091-679x(08)60963-2. [DOI] [PubMed] [Google Scholar]