Cloning and Prokaryotic Expression of Glutathione Reductase Gene in Vibrio harveyi

doi:10.13560/j.cnki.biotech.bull.1985.2015.06.029

Abstract

Abstract: The purpose of this study is to clone glutathione reductase(GR)gene from Vibrio harveyi, construct the prokaryotic expression vector of it, and obtain the corresponding expressed protein. Digested GR and pET-32a(+)were cut with the double enzymes BamH I and Xho I, then ligated with T4 ligase to construct the recombinant plasmid pET-GR. Then pET-GR was transformed into Escherichia coli BL21(DE3), which was induced by IPTG, and their expressions were analyzed by SDS-PAGE. An approximately 68.9 kD exogenous protein was observed on the SDS-PAGE. The optimal expression condition for the recombinant plasmid pET-GR was that the recombinant E. coli BL21(DE3)was induced for 4 h at 28℃ by 0. 7 mmol/L of IPTG and it expressed in E. coli as the inclusion bodies. The conclusion of the study is that GR gene from V. harveyi can efficiently express in E. coli.

Key words: Vibrio harveyi, glutathione reductase, gene cloning, prokaryotic expression

Zhu Fan, Ding Yu, Lu Yishan, Jian Jichang, Wu Zaohe. Cloning and Prokaryotic Expression of Glutathione Reductase Gene in Vibrio harveyi[J]. Biotechnology Bulletin, 2015, 31(6): 183-188.

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