重组蛋白GST-OsCAT_B的表达特性研究(英文)  

Characteristics of Recombined Protein GST-OsCAT_B Expressed in E.coli

作　　者：刘大丽[1,2,3] 鲁振强[1,3] 张革艳[1] 于婷婷[1] 王旭[1] 

机构地区：[1]省高校重点实验室生化与分子生物学、黑龙江大学生命科学学院,哈尔滨150080 [2]旨高校重点实验室甜菜遗传育种、中国农科院农作物研究院,哈尔滨150080 [3]东北林业大学森林植物生态学教育部重点实验室,哈尔滨150040

出　　处：《植物研究》2011年第6期692-695,共4页Bulletin of Botanical Research

基　　金：Supported by Heilongjiang Provincial Educational Science Foundation(12511406)

摘　　要：为了阐明水稻Catalase(CAT)的酶学功能,首先需要获得出足量的、活性的该酶蛋白。本研究克隆了水稻OsCATB基因(GenBank accession No.D26484),构建到原核表达载体pGEX-6p-3中形成重组蛋白,继而转入E.coli菌株BL21中进行表达特性研究。结果表明,GST-OsCATB融合蛋白在E.coli中进行了过量表达,表达受到诱导剂浓度、诱导时间、诱导温度和诱导体系等多因素影响;通过谷胱甘肽Sepharose-4B亲合层析,纯化出足量、活性的融合蛋白GST-OsCATB,每克表达细胞(干重)中得率为51 mg GST-OsCATB。To clarify the enzymology of catalase in rice（Oryza sativa L.）,large quantities and active of the target protein is available firstly.Here,rice OsCATB（GenBank accession No.D26484） gene was cloned and constructed with the pGEX-6p-3 vector to allow expression of OsCATB as glutathione-S-transferase（GST） fusion protein,and E.coli strain BL21 was used for expression of fusion proteins as well.The results indicated that GST-OsCATB fusion proteins were effectively expressed in E.coli,and regulated by IPTG,inducing period,temperature and others of the actions.The purified,enough and activity GST-OsCATB was obtained by affinity chromatography using glutathione-Sepharose 4B column.The final yield was 51 mg·g-1 dry cells for GST-OsCATB.

关 键 词：过氧化氢酶(CAT) 水稻 谷胱甘肽S-转移酶融合蛋白(GST) 蛋白纯化 E.COLI 

分 类 号：S511[农业科学—作物学]