2013•Chin J Naut Med & Hyperbar MedRequires access
Construction and induced expression of alkB expression vector in Escherichia coli
Min Sun, Qun Luo, Ying He, Dengyong Hou, Qingrong Wang, Xianrong Shen
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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.
Key words:
Alkane hydroxylase; Expression vector; Induced expression