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大肠杆菌GBP的定点突变与制备方法研究 被引量：1

Site-directed Mutagenesis and Methods in the Preparation of GBP in Escherichia coli
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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。 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 coll. 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.
作者 刘涛 班睿
机构地区 天津大学化工学院生物工程系
出处 《化学与生物工程》 CAS 2007年第9期50-53,共4页 Chemistry & Bioengineering
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 galactose/glucose binding protein （GBP） site-directed mutagenesis gene engineering protein purification Escherichia coli
分类号 Q319.33 [生物学—遗传学]
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参考文献13

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同被引文献14

  • 1黄显,栗大超,徐可欣,于海霞,黄福祥,胡小唐.基于微型表面等离子共振生物传感器的葡萄糖浓度测量技术[J].传感技术学报,2006,19(05B):2072-2076. 被引量：6
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  • 9Lain W W, Chu L H, Wong C L, et al. A surface plasmon resonance system for the measurement of glucose in aque- ous solution [J]. Sensors and Actuators B: Chemical, 2005, 105(2): 138-143.
  • 10Hsieh H V, Pfeiffer Z A, Amiss T J, et al. Direct detection of glucose by surface plasmon resonance with bacterial glu- cose/galactose-binding protein [J]. Biosensors and Bioelec- tronics, 2004, 19 (7): 653-660.

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化学与生物工程
2007年 第9期

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