簇毛麦谷胱甘肽硫转移酶基因HvGSTF的原核表达与酶活性鉴定  

Prokaryotic expression of a glutathione S-transferase gene HvGSTF from Haynaldia villosa and the enzyme activity

作　　者：蒋正宁[1] 赵仁慧[1] 吴旭江[1] 别同德[1] 高德荣[1] 张伯桥[1,2] 

机构地区：[1]江苏里下河地区农业科学研究所/农业部长江中下游小麦生物与遗传改良重点实验室,江苏扬州225007 [2]江苏里下河地区农业科学研究所/现代作物生产协同创新中心,江苏扬州225007

出　　处：《江苏农业学报》2016年第6期1219-1222,共4页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省自然科学青年基金项目(BK20140500);江苏省农业科技自主创新资金项目[CX(13)2022]

摘　　要：簇毛麦谷胱甘肽硫转移酶基因(Hv GSTF)是1个受白粉病诱导增强表达基因,本研究将其构入原核表达载体p ET-28a,在大肠杆菌中表达后,分离纯化重组蛋白Hv GSTF,并对其进行活性鉴定。重组蛋白Hv GSTF的分子量为29 200,浓度为0.84 mg/ml。重组蛋白Hv GSTF具有谷胱甘肽硫转移酶(GST)活性和谷胱甘肽过氧化物酶(GPX)双重活性,酶活力分别为(5.32±0.22)U/mg和(20.54±0.42)m U/mg。Hv GSTF可能参与了簇毛麦与白粉病互作,但是否参与了簇毛麦受白粉菌侵染后产生的活性氧的清除与调节仍需进一步验证。HvGSTF, a Ф -class glutathione S-transferase gene from Haynaldia villosa, is induced for expression by Blumeria graminis f. sp. tritici infection. In this study, HvGSTF gene was constructed into the vector pET-28a. After expres-sion in Escherichia coli,recombinant protein HvGSTF was purified and detected for enzyme activity. The molecular weight of recombinant HvGSTF was 29 200,and the protein concentration was 0. 84 mg/ml. Enzyme assays revealed that HvGSTF was a glutathione S-transferase （ GST） with the activity of glutathione reductase （ GPX） . The activities of GST and GPX were （5. 32±0. 22） U/mg and （20. 54±0. 42） mU/mg, respectively. The results imply that HvGSTF might have participated in the interaction between Haynaldia villosa and B. graminis f. sp. tritici because of its dual activities of both GST and GPX. It is unclear whether HvGSTF might have played a role in scavenging reactive oxygen species after infection.

关 键 词：簇毛麦 谷胱甘肽硫转移酶 HvGSTF基因 谷胱甘肽过氧化物酶 活性氧 

分 类 号：Q786[生物学—分子生物学]