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

The his7(+) gene of the fission yeast Schizosaccharomyces pombe was cloned by complementation of the recessive mutant allele his7-366. The his7(+) gene is able to complement a mutation of the Escherichia coli his1 gene, suggesting that his7(+) encodes a phosphoribosyl AMP cyclohydrase. Subcloning ex...

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Veröffentlicht in:Current genetics 1993-12, Vol.24 (6), p.491-495
Hauptverfasser: Apolinario, E. (Boston College, Chestnut Hill, MA (USA). Dept. of Biology), Nocero, M, Jin, M, Hoffman, C.S
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Sprache:eng
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Zusammenfassung:The his7(+) gene of the fission yeast Schizosaccharomyces pombe was cloned by complementation of the recessive mutant allele his7-366. The his7(+) gene is able to complement a mutation of the Escherichia coli his1 gene, suggesting that his7(+) encodes a phosphoribosyl AMP cyclohydrase. Subcloning experiments localize the gene to a 1.9-kb Xba1-Bgl2 fragment. The construction of plasmids was described 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.
ISSN:0172-8083
1432-0983
DOI:10.1007/bf00351711