| 单增李斯特氏菌ActA的原核表达与纯化 |
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| 引用本文： | 张辉,王兴龙. 单增李斯特氏菌ActA的原核表达与纯化[J]. 现代畜牧兽医, 2008, 0(3): 50-53 |
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| 作者姓名： | 张辉 王兴龙 |
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| 作者单位： | 1. 吉林大学农学部畜牧兽医学院,吉林,长春,130062,军事医学科学院军事兽医研究所,吉林,长春,130062
2. 军事医学科学院军事兽医研究所,吉林,长春,130062 |
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| 摘 要： | PCR扩增单增李斯特氏菌ActA基因,纯化后克隆到pMD 18T simple vector 中,以BamHI和EcoRI双酶切克隆载体pMD-18T/ActA,再将其亚克隆到表达载体pGEX-3X中,获得重组质粒 pGEX-3X/ActA,转化E.coli BL21(DE3)细胞,IPTG诱导融合蛋白(GST-ActA)表达,Western blot分析表明该蛋白可与单增李斯特氏菌多克隆抗体发生特异性反应.试验采用Glutathione Sepharose 4B亲合层析纯化融合蛋白.对单增李斯特氏菌ActA的原垓表达与纯化的研究为单增李斯特氏菌诊断试剂的研制及Acth的免疫原性研究奠定了基础.
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| 关 键 词： | 单增李斯特氏菌 原核表达 蛋白纯化 |
| 文章编号： | 1672-9692(2008)03-0050-04 |
| 修稿时间： | 2008-01-11 |
Expression of Listeria monocytogens ActA in Escherichia coli and Purification |
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| Zhang Hui,Wang Xinglong. Expression of Listeria monocytogens ActA in Escherichia coli and Purification[J]. Modern JOurnal of Animal Husbandry and Veterinary Medicine, 2008, 0(3): 50-53 |
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| Authors: | Zhang Hui Wang Xinglong |
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| Abstract: | The gene encoding ActA was amplified from Listeria monocytogens by PCR.The purified PCR product was cloned into pMD 18T simple vector.The pMD-18T/ActA was digested by BamHI and EcoRI double enzymes and the actA gene was subcloned into the expression vector pGEX-3X.The recombinant plasmid pGEX-3X/ActA was transformed into E.coli BL21(DE3).The bacteria was induced by IPTG and its lysate was analyzed by SDS-PAGE and Western blot.The fusion protein can react with against Listeria monocytogens polyclonal antibody.The GST-tagged protein was purififed by affinity chromatograph.This work provides a foundation for further studies on preparation for diagnosis agent and the immunogenicity of ActA. |
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| Keywords: | ActA |
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