Current Genetics

, Volume 49, Issue 6, pp 414–420 | Cite as

Cloning the Schizosaccharomyces pombe lys2 + gene and construction of new molecular genetic tools

  • Richard L. Hoffman
  • Charles S. Hoffman
Technical Note


Molecular genetic analyses in Schizosaccharomyces pombe rely on selectable markers that are used in cloning vectors or to mark targeted gene deletions and other integrated constructs. In this study, we used genetic mapping data and genomic sequence information to predict the identity of the S. pombe lys2 + gene, which is homologous to Saccharomyces cerevisiae LYS4 +. We confirmed this prediction, showing that the cloned SPAC343.16 gene can complement a lys2-97 mutant allele, and constructed the lys2 +-based cloning vector pRH3. In addition, we deleted the S. pombe his7 + gene with a lys2 +-marked polymerase chain reaction (PCR) product and the S. pombe lys2 + gene with a his7 +-marked PCR product. Strains carrying these deletions of lys2 + or his7 + serve as relatively efficient hosts for the deletion of the ade6 + gene by lys2 +- or his7 +-marked PCR products when compared with hosts carrying lys2 or his7 point mutations. Therefore, these studies provide plasmids and strains allowing the use of lys2 + as a selectable marker, along with improved strains for the use of his7 + to mark gene deletions.


Schizosaccharomyces pombe Fission yeast his7+ lys2+ Selectable marker Deletion 



We thank Val Wood for pointing out other efforts to predict the identities of S. pombe genes based on genetic and map distance information. We thank Doug Ivey for critical reading of the manuscript. This work was supported by National Institutes of Health grant GM46226 to C.S.H. and a Boston College Undergraduate Research Fellowship to R.L.H.


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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  1. 1.Biology DepartmentBoston CollegeChestnut HillUSA

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