|
|||||
|
|
| 地衣芽胞杆菌碱性蛋白酶基因克隆与表达特性 | |
| 引用本文： | 许发芝,李吕木,徐子伟,张志才.地衣芽胞杆菌碱性蛋白酶基因克隆与表达特性[J].中国微生态学杂志,2011,23(7):597-599,604. |
| 作者姓名： | 许发芝 李吕木 徐子伟 张志才 |
| 作者单位： | 1. 安徽农业大学动物科技学院,安徽,合肥,230036 2. 浙江省农业科学院,浙江,杭州,310021 3. 江苏大学食品与生物工程学院,江苏,镇江,212013 |
| 基金项目： | 安徽省教育厅自然科学基金项目(KJ2011A107); 长三角联合攻关项目(10140702021)资助 |
| 摘 要： | 目的建立高产量和高活力的地衣芽胞杆菌碱性蛋白酶基因表达体系。方法采用PCR技术克隆获得目的基因,将其连入表达质粒pET-32 a构建原核表达重组质粒,经测序鉴定后,转化BL21大肠埃希菌,不同温度下IPTG诱导表达融合蛋白,测定酶活;进一步对该基因和编码蛋白进行同源性比较和酶学性质分析。结果碱性蛋白酶基因序列全长1 149 bp,编码382个氨基酸,同源性为99%,融合蛋白分子质量为62 kD,蛋白酶酶活为29 000 U/mL,并且在25℃时是以可溶蛋白形式表达,37℃时部分蛋白以包涵体形式存在。结论此种表达体系可以成功表达具有生物活性的碱性蛋白酶,诱导温度对蛋白酶存在形式具有较大影响。 |
| 关 键 词： | 地衣芽胞杆菌 碱性蛋白酶基因 克隆 表达特性 |
Gene cloning and expression properties of Bacillus licheniformis alkaline protease |
|
| XU Fa-zhi,LI Lv-mu,XU Zi-wei,ZHANG Zhi-cai.Gene cloning and expression properties of Bacillus licheniformis alkaline protease[J].Chinese Journal of Microecology,2011,23(7):597-599,604. | |
| Authors: | XU Fa-zhi LI Lv-mu XU Zi-wei ZHANG Zhi-cai |
| Institution: | XU Fa-zhi1,LI Lv-mu1,XU Zi-wei2,ZHANG Zhi-cai3(1.College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,China,2.Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,3.School of Food and Biological Engineering,Jiangsu University,Zhenjiang 212013,China) |
| Abstract: | Objective To establish the expression system of high yield and high activity of Bacillus licheniformis alkaline protease gene.Method Bacillus licheniformis alkaline protease gene was amplified by PCR and ligated to the vector pET-32a.After identification by sequencing,the recombinant protein was expressed in E.coli BL21 though IPTG in different temperature and enzyme activity was determined.The homology of this gene and code protein was compared,enzymology characteristics of which were analyzed.Result The s... |
| Keywords: | Bacillus licheniformis Alkaline protease gene Cloning Expression properties |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |