Cloning and Expression Analysis of One Gamma-Glutamylcysteine Synthetase Gene (Hb gamma-ECS1) in Latex Production in Hevea brasiliensis
文献类型： 外文期刊
作者： Fang, Wei 1 ; Qiao, Luo Shi 1 ; Ming, Wu 1 ; Jian, Qiu 1 ; Feng, Yang Wen 1 ; Hua, Gao Hong 1 ; Zhou, Xiao Xian 1 ;
作者机构： 1.Chinese Acad Trop Agr Sci, Rubber Res Inst, Minist Agr, Key Lab Biol & Genet Resources Rubber Tree, Danzhou 571737, Hainan, Peoples R China
期刊名称：BIOMED RESEARCH INTERNATIONAL （ 影响因子：3.411； 五年影响因子：3.62 ）
ISSN： 2314-6133
年卷期： 2016 年
页码：
收录情况： SCI
摘要： Rubber tree is a major commercial source of natural rubber. Latex coagulation is delayed by thiols, which belong to the important type of antioxidants in laticifer submembrane, and is composed of glutathione (GSH), cysteine, and methionine. The rate-limiting enzyme, gamma-ECS, plays an important role in regulating the biosynthesis of glutathione under any environment conditions. To understand the relation between gamma-ECS and thiols and to correlate latex flow with one-time tapping and continuous tapping, we cloned and derived the full length of one gamma-ECS from rubber tree latex (Hb gamma-ECS1). According to qPCR analysis, the expression levels of Hb gamma-ECS1 were induced by tapping and Ethrel stimulation, and the expression was related to thiols content in the latex. Continuous tapping induced injury, and the expression of Hb gamma ECS1 increased with routine tapping and Ethrel-stimulation tapping ( more intensive tapping). According to expression in long-term flowing latex, the gene was related to the duration of latex flow. Hb gamma ECS1 was expressed in E. coli Rosetta using pET-sumo as an expression vector and the recombinant enzyme was purified; then we achieved 0.827 U/mg specific activity and about 66 kDa molecular weight. The present study can help us understand the complex role of Hb gamma-ECS in thiols biosynthesis, which is influenced by tapping.
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