| 大肠杆菌GBP的定点突变与制备方法研究 |
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| 引用本文： | 刘涛,班睿. 大肠杆菌GBP的定点突变与制备方法研究[J]. 化学与生物工程, 2007, 24(9): 50-53 |
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| 作者姓名： | 刘涛 班睿 |
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| 作者单位： | 天津大学化工学院生物工程系,天津,300072;天津大学化工学院生物工程系,天津,300072 |
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| 摘 要： | 在mglB基因上进行了E149C、 A213S、 L238S三个定点突变,并利用大肠杆菌BL21(DE3)/pET28c系统,构建了表达突变型GBP的基因工程菌BL21(DE3)/pLE3.工程菌经诱导培养后收获细胞,利用渗透休克法提取周质空间蛋白,经Ni-NTA柱纯化,从1 L培养液中可得到约3.5 mg SDS-PAGE纯度的GBP.
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| 关 键 词： | 半乳糖/葡萄糖结合蛋白 定点突变 基因工程 蛋白质纯化 大肠杆菌 |
| 文章编号： | 1672-5425（2007）09-0050-04 |
| 修稿时间： | 2007-05-18 |
Site-directed Mutagenesis and Methods in the Preparation of GBP in Escherichia coli |
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| LIU Tao,BAN Rui. Site-directed Mutagenesis and Methods in the Preparation of GBP in Escherichia coli[J]. Chemistry & Bioengineering, 2007, 24(9): 50-53 |
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| Authors: | LIU Tao BAN Rui |
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| Affiliation: | Department of Biochemical Engineering, College of Chemical Engineering, Tianjin University, Tianjin 300072, China |
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| Abstract: | In this paper,site-directed mutagenesis of E149C,A213S and L238S were carried out on gene mglB and the recombinant BL21(DE3)/pLE3 was constructed to express the GBP mutant based on expression system BL21(DE3)/ pET28c of Escherichia coli.After inducible expression and harvest cells of genetically engineered microbes,periplasmic proteins were extracted by osmotic shock method and then purified utilizing Ni-NTA.The result of SDS-PAGE showed that single protein was obtained.The mass of GBP was about 3.5 mg obtained from 1 L culture medium. |
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| Keywords: | galactose/glucose binding protein(GBP) site-directed mutagenesis gene engineering protein purification Escherichia coli |
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