1992年9月
DIRECTED MUTAGENESIS IN AN ASPOROGENOUS METHYLOTROPHIC YEAST - CLONING, SEQUENCING, AND ONE-STEP GENE DISRUPTION OF THE 3-ISOPROPYLMALATE DEHYDROGENASE GENE (LEU2) OF CANDIDA-BOIDINII TO DERIVE DOUBLY AUXOTROPHIC MARKER STRAINS
JOURNAL OF BACTERIOLOGY
- ,
- 巻
- 174
- 号
- 18
- 開始ページ
- 5988
- 終了ページ
- 5993
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- 出版者・発行元
- AMER SOC MICROBIOLOGY
A model system for one-step gene disruption for an asporogenous methylotrophic yeast, Candida boidinii, is described. In this system, the 3-isopropylmalate dehydrogenase gene (C boidinii LEU2) was selected as the target gene for disruption to derive new host strains for transformation. First, the C boidinii LEU2 gene was cloned, and its complete nucleotide sequence was determined. Next, the LEU2 disruption vectors, which had the C boidinii URA3 gene as the selectable marker, were constructed. Of the Ura+ transformants obtained with these plasmids, more than half showed a Leu- phenotype. Finally, the double-marker strains of C boidinii were derived. When vectors with repeated flanking sequences of the C. boidinii URA3 gene were used for gene disruption, Leu- Ura+ transformants changed spontaneously to a Leu- Ura- phenotype ca. 100 times more frequently than they did when plasmids without the repeated sequences were used. Southern analysis showed that these events included a one-step gene disruption and a subsequent popping out of the C boidinii URA3 sequence from the transformant chromosome.
- リンク情報
- ID情報
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- ISSN : 0021-9193
- eISSN : 1098-5530
- Web of Science ID : WOS:A1992JN26400029