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Current Genetics

, Volume 24, Issue 6, pp 491–495 | Cite as

Cloning and manipulation of the Schizosaccharomyces pombe his7+gene as a new selectable marker for molecular genetic studies

  • Ethel Apolinario
  • Mary Nocero
  • Mei Jin
  • Charles S. Hoffman
Original Articles

Abstract

We have cloned the his7+ gene of the fission yeast Schizosaccharomyces pombe by complementation of the recessive mutant allele his7-366. The his7+ gene is able to complement a mutation of the Escherichia coli hisI gene, suggesting that his7+ encodes a phosphoribosyl-AMP cyclohydrase. Subcloning experiments localize the gene to a 1.9-kb XbaI-BglII fragment. We describe the construction of plasmids to facilitate the use of his7+ as a selectable marker in S. pombe studies. Plasmid pEA2 carries his7+ cloned into the pUC18 polylinker. From either pEA2 or the original his7+ clone, pMN1, fragments carrying his7+ can be isolated using a variety of restriction enzymes for the construction of gene disruptions. Plasmid pEA500 is a cloning vector that carries his7+ and ars1, yet retains the ability to use the blue/white color screen to identify recombinants.

Key words

Fission yeast Selectable marker Gene disruptions Cloning vector 

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Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Ethel Apolinario
    • 1
  • Mary Nocero
    • 1
  • Mei Jin
    • 1
  • Charles S. Hoffman
    • 1
  1. 1.Department of BiologyBoston CollegeChestnut HillUSA

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