| 猪链球菌2型epf基因的原核表达及其蛋白纯化 |
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| 引用本文： | 葛长城,王海丽,徐公义.猪链球菌2型epf基因的原核表达及其蛋白纯化[J].安徽农业科学,2009,37(15):6916-6917. |
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| 作者姓名： | 葛长城 王海丽 徐公义 |
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| 作者单位： | 1. 聊城职业技术学院,山东,聊城,252000;聊城绿亚禽业有限公司,山东,聊城,252000
2. 聊城职业技术学院,山东,聊城,252000 |
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| 基金项目： | 山东省教育厅科技计划项目 |
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| 摘 要： | 为了克隆、表达SS2菌株轲基因片段，分析蛋白活性，根据GenBank SS2 epf基因序列设计引物，克隆epf基因片段并进行序列分析，构建原核表达质粒pGEX4T-2-epf，在大肠杆菌中诱导带有GST标签的融合蛋白rEF-GST的表达；亲和层析法纯化融合蛋白rEF-GST，用凝血酶切除重组蛋白中的GST，获得纯化的EF抗原。经Westernblot鉴定，EF可与SS2多克隆抗血清发生特异性反应。
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| 关 键 词： | 猪链球菌2型 epf基因 胞外因子 原核表达 蛋白纯化 |
Prokaryotic Expression of epf Gene of Streptococcus suis 2 and Its Protein Purification |
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| Institution: | GE Chang-cheng et al (Liaecheng Vocational and Technical College, Liaocheng, Shandong 252000) |
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| Abstract: | To clone and express the truncated epf gene of SS2 strain.epf gene fragment was amplified by PCR from and cloned into prokaryotic expression plasmid pGEX4T-2 to form pGEX4T-2-epf.The expression of recombinant protein with GST was induced in E.coli TG1 and the fusion protein(rEF-GST) was purified by affinity chromatography,and then GST was cut with thrombin protease.The expressed and purified proteins were analyzed by Western blot.The purified EF could be recognized by ployclonal anti-SS2 antibody. |
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| Keywords: | Streptococcus suis 2 epf gene Extracellular factor Prokaryotic expression Protein purification |
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