Plasmid recombination intermediates generated in a Saccharomyces cerevisiae cell-free recombination system
- PMID: 3915541
- PMCID: PMC366963
- DOI: 10.1128/mcb.5.9.2361-2368.1985
Plasmid recombination intermediates generated in a Saccharomyces cerevisiae cell-free recombination system
Abstract
We have developed an assay utilizing Saccharomyces cerevisiae cell extracts to catalyze recombination in vitro between homologous plasmids containing different mutant alleles of the tet gene. Electrophoretic analysis of product DNA indicated that a number of novel DNA species were formed during the reaction. These species migrated through agarose gels as distinct bands with decreased electrophoretic mobility compared with the substrate DNA. The DNA from each individual band was purified and shown to be enriched 5- to 100-fold for tetracycline-resistant recombinants by using a transformation assay. The structure of the DNA molecules present in these bands was determined by electron microscopy. Recombination between circular substrates appeared to involve the formation and processing of figure-eight molecules, while recombination between circular and linear substrates involved the formation of molecules in which a circular monomer had a monomer-length linear tail attached at a region of homology.
Similar articles
-
Genetic recombination of homologous plasmids catalyzed by cell-free extracts of Saccharomyces cerevisiae.Cell. 1983 Dec;35(3 Pt 2):805-13. doi: 10.1016/0092-8674(83)90113-7. Cell. 1983. PMID: 6360380
-
Transformation-associated recombination between diverged and homologous DNA repeats is induced by strand breaks.Yeast. 1994 Jan;10(1):93-104. doi: 10.1002/yea.320100109. Yeast. 1994. PMID: 8203155
-
Induction of recombination between homologous and diverged DNAs by double-strand gaps and breaks and role of mismatch repair.Mol Cell Biol. 1994 Jul;14(7):4802-14. doi: 10.1128/mcb.14.7.4802-4814.1994. Mol Cell Biol. 1994. PMID: 8007979 Free PMC article.
-
Copy number amplification of the 2 micron circle plasmid of Saccharomyces cerevisiae.J Theor Biol. 1986 Mar 21;119(2):197-204. doi: 10.1016/s0022-5193(86)80074-1. J Theor Biol. 1986. PMID: 3525993
-
The 2 micron circle plasmid of Saccharomyces cerevisiae.Yeast. 1988 Mar;4(1):27-40. doi: 10.1002/yea.320040104. Yeast. 1988. PMID: 3059711 Review. No abstract available.
Cited by
-
Homologous recombination in procaryotes.Microbiol Rev. 1988 Mar;52(1):1-28. doi: 10.1128/mr.52.1.1-28.1988. Microbiol Rev. 1988. PMID: 3280962 Free PMC article. Review. No abstract available.
-
Partial purification of an enzyme from Saccharomyces cerevisiae that cleaves Holliday junctions.Proc Natl Acad Sci U S A. 1985 Nov;82(21):7247-51. doi: 10.1073/pnas.82.21.7247. Proc Natl Acad Sci U S A. 1985. PMID: 3903750 Free PMC article.
-
Double-strand-break repair and recombination catalyzed by a nuclear extract of Saccharomyces cerevisiae.EMBO J. 1991 Apr;10(4):987-96. doi: 10.1002/j.1460-2075.1991.tb08033.x. EMBO J. 1991. PMID: 2009864 Free PMC article.
-
Joining of nonhomologous DNA double strand breaks in vitro.Nucleic Acids Res. 1988 Feb 11;16(3):907-24. doi: 10.1093/nar/16.3.907. Nucleic Acids Res. 1988. PMID: 3344222 Free PMC article.
-
Structural effect of donor DNA on the initiation of recombination for double strand break repair in human nuclear extracts.Nucleic Acids Res. 1992 Oct 11;20(19):5167-72. doi: 10.1093/nar/20.19.5167. Nucleic Acids Res. 1992. PMID: 1408832 Free PMC article.
References
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
