• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of geneticsGeneticsCurrent IssueInformation for AuthorsEditorial BoardSubscribeSubmit a Manuscript
Genetics. Mar 1994; 136(3): 849–856.
PMCID: PMC1205890

Efficient Targeted Integration at Leu1-32 and Ura4-294 in Schizosaccharomyces Pombe

Abstract

Homologous integration into the fission yeast Schizosaccharomyces pombe has not been well characterized. In this study, we have examined integration of plasmids carrying the leu1(+) and ura4(+) genes into their chromosomal loci. Genomic DNA blot analysis demonstrated that the majority of transformants have one or more copies of the plasmid vector integrated via homologous recombination with a much smaller fraction of gene conversion to leu1(+) or ura4(+). Non-homologous recombination events were not observed for either gene. We describe the construction of generally useful leu1(+) and ura4(+) plasmids for targeted integration at the leu1-32 and ura4-294 loci of S. pombe.

Full Text

The Full Text of this article is available as a PDF (2.9M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Carr AM, MacNeill SA, Hayles J, Nurse P. Molecular cloning and sequence analysis of mutant alleles of the fission yeast cdc2 protein kinase gene: implications for cdc2+ protein structure and function. Mol Gen Genet. 1989 Jul;218(1):41–49. [PubMed]
  • Feinberg AP, Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. [PubMed]
  • Grallert B, Nurse P, Patterson TE. A study of integrative transformation in Schizosaccharomyces pombe. Mol Gen Genet. 1993 Apr;238(1-2):26–32. [PubMed]
  • Grimm C, Kohli J, Murray J, Maundrell K. Genetic engineering of Schizosaccharomyces pombe: a system for gene disruption and replacement using the ura4 gene as a selectable marker. Mol Gen Genet. 1988 Dec;215(1):81–86. [PubMed]
  • Hayles J, Aves S, Nurse P. suc1 is an essential gene involved in both the cell cycle and growth in fission yeast. EMBO J. 1986 Dec 1;5(12):3373–3379. [PMC free article] [PubMed]
  • Hill J, Donald KA, Griffiths DE, Donald G. DMSO-enhanced whole cell yeast transformation. Nucleic Acids Res. 1991 Oct 25;19(20):5791–5791. [PMC free article] [PubMed]
  • Kikuchi Y, Kitazawa Y, Shimatake H, Yamamoto M. The primary structure of the leu1+ gene of Schizosaccharomyces pombe. Curr Genet. 1988 Oct;14(4):375–379. [PubMed]
  • Levin HL, Weaver DC, Boeke JD. Two related families of retrotransposons from Schizosaccharomyces pombe. Mol Cell Biol. 1990 Dec;10(12):6791–6798. [PMC free article] [PubMed]
  • Russell P, Nurse P. cdc25+ functions as an inducer in the mitotic control of fission yeast. Cell. 1986 Apr 11;45(1):145–153. [PubMed]
  • Russell P, Nurse P. Negative regulation of mitosis by wee1+, a gene encoding a protein kinase homolog. Cell. 1987 May 22;49(4):559–567. [PubMed]
  • Sikorski RS, Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. [PMC free article] [PubMed]

Articles from Genetics are provided here courtesy of Genetics Society of America

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...