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Construction of protease-deficient Candida boidinii strains useful for recombinant protein production:: Cloning and disruption of proteinase A gene (PEP4) and proteinase B gene (PRB1)
被引：17
|作者：
Komeda, T
Sakai, Y
Kato, N
Kondo, K
机构：
[1] Kirin Brewery Co Ltd, Cent Labs Key Technol, Kanagawa Ku, Yokohama, Kanagawa 2360004, Japan
[2] Kyoto Univ, Grad Sch Agr, Div Appl Life Sci, Sakyo Ku, Kyoto 6068502, Japan
关键词：
Candida boidinii;
proteinase A;
PEP4;
proteinase B;
PRB1;
D O I：
10.1271/bbb.66.628
中图分类号：
Q5 [生物化学];
Q7 [分子生物学];
学科分类号：
071010 ;
081704 ;
摘要：
The yeast Candida boidinii PEP4 and PRB1 genes, encoding proteinase A (PrA) and proteinase B (PrB), respectively, have been cloned and their primary structures were analyzed. The open reading frames of the PEP4 gene (1263 bp encoding a protein of 420 amino acids) and the PRB1 gene (1683 bp encoding a protein of 560 amino acids) were found. The deduced amino acid sequences of PrA and PrB are very similar to Saccharomyces cereviside PrA and PrB (64% and 61% identities, respectively). Both PEP4 and PRB1 genes were disrupted in the C. boidinii genome by one-step gene disruption. The resultant pep4Delta and the pep4Delta prb1Delta strains lost protease activity when compared with the wild-type original strain. The constructed C. boidinii strains are expected to be useful hosts for heterologous protein production.
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页码：628 / 631
页数：4
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