| 角蛋白酶基因kerB的提取、 克隆及表达 |
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| 引用本文： | 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].吉林大学学报(理学版),2003,41(3):365-368. |
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| 作者姓名： | 梁斌 邢述 付学奇 林祥 |
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| 作者单位： | 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 |
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| 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.
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| 关 键 词： | 角蛋白酶 地衣芽孢杆菌L-25 kerB基因 |
| 文章编号： | 1671-5489(2003)03-0365-04 |
| 收稿时间： | 2003-01-22 |
| 修稿时间： | 2003年1月22日 |
Extraction, Cloning and Expression of Keratinase Gene kerB |
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| LIANG Bin,XING Shu,FU Xue qi.Extraction, Cloning and Expression of Keratinase Gene kerB[J].Journal of Jilin University: Sci Ed,2003,41(3):365-368. |
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| Authors: | LIANG Bin XING Shu FU Xue qi |
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| Institution: | 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 |
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| Abstract: | A 1.4 kb keratinase gene( ker B) was isolated from Bacillus licheniformis 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 PCR2.1.Then the keratinase gene within PCR2.1 was excised by restriction endonuclease XbaI SpeI , and inserted into the plasmid of PUB18 P43 for expressing. This new plasmid(PLK18) was transformed into protease deficient strain Bacillus subtilis DB104 competent cells. The FD 8(DB104/PLK18) strain could grow rapidly on feather medium and completely hydrolyze all feathers in the medium. |
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| Keywords: | keartinase Bacillus licheniformis L-25 strain kerB gene |
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