Establishment of a screening system for essential genes from the pathogenic yeast Candida glabrata: identification of a putative TEM1 homologue

Authors

  • Y. Miyakawa,

    1. Division of Biotechnology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Takeda, Kofu, Yamanashi, Japan
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  • T. Hara,

    1. Division of Biotechnology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Takeda, Kofu, Yamanashi, Japan
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  • Y. Iimura

    1. Division of Biotechnology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Takeda, Kofu, Yamanashi, Japan
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Yozo Miyakawa, Division of Biotechnology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 4-3-11 Takeda, Kofu, Yamanashi 400-8511, Japan. E-Mail : ymiya@yamanashi.ac.jp

Abstract

Aims:  To establish a system for screening and identification of essential genes from the pathogenic haploid yeast Candida glabrata by using temperature-sensitive (ts) mutants.

Methods and Results:  Based on the general concepts that ts mutations are generated within essential genes in the genome by virtue of point mutation, we attempted to establish a system where essential genes were screened and identified from the C. glabrata genomic DNA library by the complementation of ts point mutations. By using this system, we successfully identified a putative TEM1 homologue as an essential gene by the complementation of a point mutation (-GAT-/-AAT- corresponding to Asp-143/Asn substitution) within its coding region in a ts mutant, T-3.

Conclusions:  We were able to establish a system for screening and identification of the essential genes, such as the TEM1 homologue, from the pathogenic yeast C. glabrata, as the gene that complements ts mutation.

Significance and Impact of the Study:  The identification of essential genes, by using the present system, may provide novel potential antifungal targets.

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