茶树花青素还原酶基因在大肠杆菌中的表达及优化

doi:10.13305/j.cnki.jts.2011.04.014

Abstract

Abstract: Anthocyanin reductase is a key enzyme in the biosynthesis of EC and EGC in tea plant. The open reading frame of anthocyanin reductase gene (ANR), which encoding a 337 amino acids protein, was cloned from tea plant 〔Camellia sinensis (L.) O. Kuntze 〕 by RT-PCR. The deduced protein molecular weight was 37kD and its theoretical isoelectric point was 6.54. The gene was cloned into the expression vector pET32a (+) for expression in prokaryotic cells. The SDS-PAGE results showed that the anthocyanin reductase peoteins was expressed in Escherichia coli rosetta. The optimal inducing conditions including time, temperature, IPTG concentration, ampicillin concentration were studied. The deduced protein was purified and its activity was detected by HPLC.

Key words: tea plant [Camellia sinensis (L.) O. Kuntze], anthocyanin reductase, prokaryotic expression, enzyme activity

CLC Number:

S571.1
Q554

LUO Yang, WANG Hong-xue, WANG Yun-sheng, LU Zhong-wei, LIU Ya-jun, SHAN Yu, CHEN Xiao-tian, YE Hui, GAO Li-ping, XIA Tao. Expression and Optimization of Anthocyanin Reductase Gene of Tea Plant [Camellia sinensis (L.) O. Kuntze] in Escherichia coli[J]. Journal of Tea Science, 2011, 31(4): 326-332.

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Expression and Optimization of Anthocyanin Reductase Gene of Tea Plant [Camellia sinensis (L.) O. Kuntze] in Escherichia coli

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