Construction and induced expression of alkB expression vector in Escherichia coli
10.3760/cma.j.issn.1009-6906.2013.01.016
- VernacularTitle:烷烃羟化酶基因alkB表达载体的构建及其在大肠杆菌中的诱导表达
- Author:
Min SUN
1
;
Qun LUO
1
;
Ying HE
1
;
Deng-yong HOU
1
;
Qing-rong WANG
1
;
Xian-rong SHEN
1
Author Information
1. 海军医学研究所,上海,200433
- Publication Type:Journal Article
- Keywords:
Alkane hydroxylase;
Expression vector;
Induced expression
- From:
Chinese journal of nautical medicine and hyperbaric medicine
2013;20(1):50-53
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct alkB-expressing vector and investigate alkB protein expression in Escherichia coli.Methods The alkane hydroxylase (alkB) gene was synthesized in vitro.An alkB-expressing vector,pCom8-alkB,was constructed and then transformed into E.coli,DH5α by using CaCl2.Changes in the expression of alkB were detected by SDS-PAGE in E.coli,DH5α by induction with diesel oil.Results SDSPAGE analysis demonstrated that alkB was successfully expressed after induction.The expression levels of alkB were highest,when alkB expression was induced by adding diesel oil into the culture at the final concentration of 2％-3％ (v/v),and the expression levels increased with the increase of induction time.Conclusions The alkB-expressing vector was successfully constructed,and the regular pattern for the expression of target protein under different induction conditions was clearly explained.
