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| 碱性蛋白酶PB92在枯草芽孢杆菌中高效转化方法的建立及其分泌表达 | |
| 引用本文： | 孙同毅,邵建新,高媛媛,陈丽梅,高志芹,林伟平,路福平.碱性蛋白酶PB92在枯草芽孢杆菌中高效转化方法的建立及其分泌表达[J].江苏农业学报,2009,25(3). |
| 作者姓名： | 孙同毅 邵建新 高媛媛 陈丽梅 高志芹 林伟平 路福平 |
| 作者单位： | 1. 潍坊医学院生物技术实验室,山东潍坊,261053 2. 潍坊医学院化学分析实验室,山东潍坊,261053 3. 天津市工业微生物重点实验室,天津科技大学生物工程学院,天津300457 |
| 基金项目： | 国家高技术研究发展"863"计划，国家科技资源平台项目 |
| 摘 要： | 枯草芽孢杆菌转换效率低一直制约着该菌基因改造技术的应用,本研究比较了化学法和高渗电转化法对枯草芽孢杆菌转化率的影响,发现高渗电转化法的转化效率较高.从Bacillus alcalophillus PB92中扩增出碱性蛋白酶基因Mapr,构建成重组分泌型表达载体pWB980-Mapr,碱性蛋白酶基因在枯草芽孢杆菌DB104中得到表达.SDS-PAGE 分析,重组蛋白酶的分子量为 28 000.pWB980-Mapr在枯草芽孢杆菌中表达的酶活为1 563 U/ml. |
| 关 键 词： | 碱性蛋白酶 枯草芽孢杆菌 高渗电转化法 |
Establishment of High-efficiency Transformation and Expression of the Gene Encoding Alkaline Protease PB92 in Bacillus subtilis DB104 |
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| SUN Tong-yi,SHAO Jian-xin,GAO Yuan-yuan,CHEN Li-mei,GAO Zhi-qin,LIN Wei-ping,LU Fuping.Establishment of High-efficiency Transformation and Expression of the Gene Encoding Alkaline Protease PB92 in Bacillus subtilis DB104[J].Jiangsu Journal of Agricultural Sciences,2009,25(3). | |
| Authors: | SUN Tong-yi SHAO Jian-xin GAO Yuan-yuan CHEN Li-mei GAO Zhi-qin LIN Wei-ping LU Fuping |
| Institution: | 1.Laboratory of Biotechnology;Weifang Medical College;Weifang 261053;China;2.Laboratory of Chemical Analysis;3.Tianjin Key Laboratory of Industrial Microbiology;College of Bioengineering;Tianjin University of Science & Technology;Tianjin;300457 |
| Abstract: | The application of genetical modification technology has been constrained by low transformation efficiency in Bacillus subtilis.Compared with chemical transformation,the hypertonic electroporation exhibited higher transformation efficiency in this experiment.The sequence of alkaline protease gene Mapr cloned from Bacillus alkalophillus PB92 chromosomeal DNA was inserted into B.subtilis vector pWB980 to yield the recombinant plasmid pWB980-Mapr.After the transformaiton by hypertonic electroporation,pWB980-Ma... |
| Keywords: | alkaline protease Bacillus subtilis hypertonic electroporation |
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