一种高效表达碱性蛋白酶的新型启动子的筛选及研究

doi:10.13560/j.cnki.biotech.bull.1985.2017-0748

生物技术通报 ›› 2018, Vol. 34 ›› Issue (1): 208-214.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0748

一种高效表达碱性蛋白酶的新型启动子的筛选及研究

陈坤¹, 袁飞燕¹, 柴昊男¹, 刘焕¹, 王兴吉², 张会图¹, 路福平¹

1. 工业发酵微生物教育部重点实验室天津科技大学生物工程学院,天津 300457;
2. 山东隆科特酶制剂有限公司,沂水 276400

收稿日期:2017-09-08 出版日期:2018-01-26 发布日期:2018-01-22
作者简介:陈坤,男,硕士研究生,研究方向：应用微生物与酶工程;E-mail：chenkun2020@163.com
基金资助:
国家重点研发计划（2017YFB0308401）

Screening and Study of a New Promoter with Effective Expression of Alkaline Protease

CHEN Kun¹, YUAN Fei-yan¹, CHAI Hao-nan¹, LIU Huan¹, WANG Xing-ji², ZHANG Hui-tu¹, LU Fu-ping¹

1. Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,College of Biotechnology,Tianjin University of Science & Technology,Tianjin 300457;
2. LONGDA Biotechnology,Yishui 276400

Received:2017-09-08 Published:2018-01-26 Online:2018-01-22

摘要/Abstract

摘要： 碱性蛋白酶广泛的应用价值及市场供不应求的现状,旨在构建碱性蛋白酶高效表达系统。通过蛋白电泳和质谱检测的方法,由地衣芽孢杆菌中筛选获得一种新型启动子pChi,以两种已知强组成型启动子pShuttle-09和pyxiE为对照,研究其对克劳氏芽孢杆菌碱性蛋白酶与地衣芽孢杆菌碱性蛋白酶基因的表达活性。成功构建了6种重组表达载体及重组菌,结果表明,pChi的启动强度是pShuttle-09的1.25倍,pyxiE的2倍,从而为介导枯草芽孢杆菌表达系统中异源基因的表达奠定基础。

关键词: 碱性蛋白酶, 高效表达系统, 启动子, 异源表达

Abstract: The alkaline protease has extensive applications but in short supply,for which the efficient expression system of alkaline protease is constructed. A new type of promoter pChi was screened from the Bacillus licheniformis by SDS-PAGE and mass spectrometric detection. With the other two strong constitutive promoters pShuttle-09 and pyxiE as the control,pChi’s expression activities in the alkaline proteases from B. clausii and B. licheniformis was investigated. Further,six types of recombinant expression vectors and bacteria were constructed. The results showed that the functional expression of pChi was 1.25 times of pShuttle-09 and 2 times of pyxiE,which offers the foundation for the gene heterologous expression in mediated B. subtilis expression system.

Key words: alkaline protease, efficient expression system, promoter, heterologous expression

陈坤, 袁飞燕, 柴昊男, 刘焕, 王兴吉, 张会图, 路福平. 一种高效表达碱性蛋白酶的新型启动子的筛选及研究[J]. 生物技术通报, 2018, 34(1): 208-214.

CHEN Kun, YUAN Fei-yan, CHAI Hao-nan, LIU Huan, WANG Xing-ji, ZHANG Hui-tu, LU Fu-ping. Screening and Study of a New Promoter with Effective Expression of Alkaline Protease[J]. Biotechnology Bulletin, 2018, 34(1): 208-214.

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一种高效表达碱性蛋白酶的新型启动子的筛选及研究

Screening and Study of a New Promoter with Effective Expression of Alkaline Protease

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