| 重组蛋白GST-OsCATB的表达特性研究 |
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| 引用本文： | 刘大丽；鲁振强；张革艳；于婷婷；王旭.重组蛋白GST-OsCATB的表达特性研究[J].植物研究,2011,31(6):692-695. |
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| 作者姓名： | 刘大丽；鲁振强；张革艳；于婷婷；王旭 |
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| 作者单位： | 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.省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 |
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| 基金项目： | Supported by Heilongjiang Provincial Educational Science Foundation(12511406) |
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| 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。
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| 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 |
Characteristics of Recombined Protein GST-OsCATB Expressed in E.coli |
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| LIU Da-Li, LU Zhen-Qiang, ZHANG Ge-Yan YU Ting-Ting WANG Xu.Characteristics of Recombined Protein GST-OsCATB Expressed in E.coli[J].Bulletin of Botanical Research,2011,31(6):692-695. |
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| Authors: | LIU Da-Li LU Zhen-Qiang ZHANG Ge-Yan YU Ting-Ting WANG Xu |
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| Institution: | LIU Da-Li1,2,3 LU Zhen-Qiang1,3 ZHANG Ge-Yan1 YU Ting-Ting1 WANG Xu1(1.Key Laboratory of Molecular Biology,of Heilongjiang Provincial,College of Life Sciences,Heilongjiang University,Harbin 150080,2.Key Laboratory of Sugar Beet Genetics and Breeding,Academy of Crop Sciences,Chinese Academy of Agricultural Sciences,3.Key Laboratory of Forest Plant Ecology of Northeast Forestry University,Ministry of Education,Harbin 150040) |
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| 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 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. |
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| Keywords: | Catalase rice(Oryza sativa L ) glutathione-S-transferase(GST) fusion protein purification Escherichia coli |
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