| 牛GM-CSF与金黄色葡萄球菌黏附素FnBPB共表达质粒的构建与原核表达 |
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| 引用本文： | 范鑫,郝永清,张爱荣,陈晓杰.牛GM-CSF与金黄色葡萄球菌黏附素FnBPB共表达质粒的构建与原核表达[J].动物医学进展,2011(10):79-82. |
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| 作者姓名： | 范鑫 郝永清 张爱荣 陈晓杰 |
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| 作者单位： | 内蒙古农业大学兽医学院; |
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| 摘 要： | 采用PCR方法对牛粒细胞-巨噬细胞集落刺激因子(GM-CSF)和金黄色葡萄球菌FnBPB的D区进行特异性的扩增,并通过重叠延伸PCR扩增GM-CSF-FnBPB串联基因,构建了克隆质粒pMD19-GM-CSF-FnBPB。利用表达载体pET-32a(+)对该融合基因片段进行原核表达,SDS-PAGE分析表明,在1mmol/L IPTG诱导浓度下,在39ku处出现了与目的蛋白一致的外源蛋白带,Western blot分析表明,该蛋白具有反应原性,进而证明该融合基因成功在原核细胞中表达。
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| 关 键 词： | 金黄色葡萄球菌 粒细胞-巨噬细胞集落刺激因子 纤连素结合蛋白B 重叠延伸PCR克隆 原核表达 |
Construction and Prokaryotic Expression of the Co-expression Plasmid with FnBPB and GM-CSF Gene Encoding Staphylococcus aureus Adhesion |
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| FAN Xin,HAO Yong-qing,ZHANG Ai-rong,CHEN Xiao-jie.Construction and Prokaryotic Expression of the Co-expression Plasmid with FnBPB and GM-CSF Gene Encoding Staphylococcus aureus Adhesion[J].Progress In Veterinary Medicine,2011(10):79-82. |
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| Authors: | FAN Xin HAO Yong-qing ZHANG Ai-rong CHEN Xiao-jie |
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| Institution: | FAN Xin,HAO Yong-qing,ZHANG Ai-rong,CHEN Xiao-jie(College of Veterinary Medicine,Inner Mongolia Agricultural Universtiy,Hohhot,Inner Mongolia,010018,China) |
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| Abstract: | Granulocyte-macrophage colony stimulating factor(GM-CSF) and fibronectin-binding proteins B(FnBPB) and fibrinogen-binding(GM-CSF) were amplified by PCR from Staphylococcus aureus chromosomal DNA and the amplified sequence GM-CSF-FnBPB was cloned into pMD19-T vector by overlap extension PCR.The fusion gene was successfully coloned into expression vector pET-32a.The constructed plasmid pET-32a-GM-CSF-FnBPB was transformed into E.coli BL21(DE3) competent cell.The bactera were induced by 1 mmol/L IPTG and analy... |
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| Keywords: | Staphylococcus aureus granulocyte-macrophage colony stimulating factor fibronectin-binding proteins B overlap extension PCR prokaryotic expression |
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