角蛋白酶基因kerB的提取、 克隆及表达

角蛋白酶基因kerB的提取、克隆及表达

梁斌¹，邢述¹，付学奇¹, 林祥²

1. 吉林大学生命科学学院，长春 130023; 2. 英哥伦比亚大学农学院，温哥华 V 6T 1Z4，加拿大

收稿日期:2003-01-22 修回日期:1900-01-01 出版日期:2003-07-26 发布日期:2003-07-26
通讯作者: 付学奇

Extraction, Cloning and Expression of Keratinase Gene kerB

LIANG Bin¹， XING Shu¹， FU Xue-qi¹, LIN Xiang< sup>2

1. College of Life Science, Jilin University, Changchun 130023， China; 2. Faculty of Agricultural Science, The University of British Columbia, Vancouve r, B.C. V6T 1Z4, Canada

Received:2003-01-22 Revised:1900-01-01 Online:2003-07-26 Published:2003-07-26
Contact: FU Xue-qi

摘要/Abstract

摘要： 从角蛋白酶产生菌株地衣芽孢杆菌L-25中提取基因组DNA，借助特定引物通过聚合酶链反应(PCR)扩增得到kerB基因片段. 扩增后的kerB片段通过分子生物学方法克隆到产酶缺陷型枯草芽孢杆菌DB104菌株敏感细胞中进行表达. 结果表明，表达成功的FD-8菌株（DB104/pLK18）可以在羽毛培养基上生长并完全水解羽毛.

关键词: 角蛋白酶, 地衣芽孢杆菌L-25, kerB基因

Abstract: A 1.4 kb keratinase gene(kerB) was isolated from Bacillus licheniformi s L-25 strain by a polymerase chain reaction(PCR) with L-25 genomic DNA as the template. Amplified keratinase gene was directly cloned into a cloning vector PC R2.1. Then the keratinase gene within PCR2.1 was excised by restriction endo nuclease XbaI-SpeI, and inserted into the plasmid of PUB18-P43 for expressing. This new plasmid(PLK18) was transformed into protease-deficient strain Bacillus s ubtilis DB104 competent cells. The FD-8(DB104/PLK18) strain could grow rapidly o n feather-medium and completely hydrolyze all feathers in the medium.

Key words: keartinase, Bacillus licheniformis L-25 strain, kerB gene

中图分类号:

Q534

梁斌，邢述，付学奇, 林祥. 角蛋白酶基因kerB的提取、克隆及表达[J]. J4, 2003, 41(03): 365-368.

LIANG Bin， XING Shu， FU Xue-qi, LIN Xiang. Extraction, Cloning and Expression of Keratinase Gene kerB[J]. J4, 2003, 41(03): 365-368.