Constitutive-expression vector and its construction method and application
Title：
Constitutive-expression vector and its construction method and application
Application Number：
 200410083849
Application Date：
 2004/10/20
Announcement Date：
 2006/04/26
Pub. Date：
Publication Number：
 1763204
Announcement Number：
Grant Date：
Granted Pub. Date：
ApplicationType：
 Invention
State/Country：
 11[China|beijing]
IPC：
 C12N 15/63, C12N 15/66, C12N 15/52, C12N 9/00, C12N 15/70
Applicant(s)：
 Tsinghua University
Inventor(s)：
 Zhou Hang, Yu Huimin, Luo Hui, Shen Zhongyao
Key Words：
 Constitutive-expression vector, construction method, application
Abstract：
 The present invention is one kind of constitutive expression vector and its construction process and application in expressing GL-7-ACA acrylatase. The constitutive expression vector of the present invention is inducible plasmid vector with regulating gene eliminated or deleted. Through utilizing of pMAL-p2x, pGEX, pMBP, pKK223 and other plasmid expression vector containing tac promoter as template, designing primer to gene lac Iq ends, and PCR amplifying the whole plasmid except lac Iq gene, optimized constitutive expression vector pMALC is obtained. Through the said process, the present invention obtain thallus with dry weight of about 1 g/L, total protein amount of about 200 mg/L, enzyme accounting for 40-50 % of the total protein, expressed GL-7-ACA acrylatase amount basically the same as inducing expression amount and enzyme activity of 220 U/L. The constitutive expression vector will play important role in the production of foreign protein.
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2008-4-17
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