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作 者 ：刘大丽;;鲁振强;*;张革艳;于婷婷;王旭

期 刊 ：植物研究 2011年 31卷 6期 页码：692-695

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；

Keywords:Catalase, rice(Oryza sativa L.), glutathione-S-transferase(GST) fusion protein, purification, Escherichia coli,

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OsCAT_B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-OsCAT_B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-OsCAT_B，每克表达细胞（干重）中得率为51 mg GST-OsCAT_B。

Abstract:To clarify the enzymology of catalase in rice(Oryza sativa L.), large quantities and active of the target protein is available firstly. Here, rice OsCAT_B(GenBank accession No.D26484) gene was cloned and constructed with the pGEX-6p-3 vector to allow expression of OsCAT_B 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-OsCAT_B 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-OsCAT_B was obtained by affinity chromatography using glutathione-Sepharose 4B column. The final yield was 51 mg·g^-1 dry cells for GST-OsCAT_B.

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