| 金黄色葡萄球菌Fc段受体结合蛋白A基因的克隆与原核表达 |
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| 引用本文： | 柏家林,白喜龙,张从党,李向茸,冯若飞,马忠仁,蔡葵蒸. 金黄色葡萄球菌Fc段受体结合蛋白A基因的克隆与原核表达[J]. 中国兽医科学, 2012, 0(5): 505-510 |
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| 作者姓名： | 柏家林 白喜龙 张从党 李向茸 冯若飞 马忠仁 蔡葵蒸 |
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| 作者单位： | 甘肃省动物细胞工程技术研究中心;西北民族大学生命科学与工程学院 |
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| 基金项目： | 甘肃省兰州市生物技术专项(2010-1-1-159) |
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| 摘 要： | 为研制葡萄球菌蛋白A(staphylococcal protein A,SPA)的相关免疫试剂盒,设计了1对引物,从金黄色葡萄球菌基因组中扩增出了876bp的细胞壁结合蛋白SPA基因,该基因编码292个氨基酸,包含了E、D、A、B、C 5个IgG结合结构域。构建了原核表达载体pET28a(+)-SPA,经IPTG诱导,表达出分子质量约35.0ku的SPA蛋白,并应用亲和层析柱纯化了SPA蛋白,Western-blot分析表明纯化的SPA蛋白具有生物学活性。
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| 关 键 词： | 金黄色葡萄球菌蛋白A 基因克隆 原核表达 亲和层析 |
Cloning and prokaryotic expression of Fc-binding protein A gene of Staphylococcal aureus |
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| BAI Jia-lin,BAI Xi-long,ZHANG Cong-dang,LI Xiang-rong,FENG Ruo-fei,MA Zhong-ren,CAI Kui-zheng. Cloning and prokaryotic expression of Fc-binding protein A gene of Staphylococcal aureus[J]. Chinese Veterinary Science, 2012, 0(5): 505-510 |
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| Authors: | BAI Jia-lin BAI Xi-long ZHANG Cong-dang LI Xiang-rong FENG Ruo-fei MA Zhong-ren CAI Kui-zheng |
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| Affiliation: | 1.Gansu Engineering Research Center for Animal Cell,Lanzhou 730030,China;2.College of Life Science and Engineering,Northwest University for Nationalities,Lanzhou 730030,China) |
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| Abstract: | To develop staphylococcal protein A(SPA) relative immunological kits,a pair of primers for SPA was designed according to the published genomic DNA sequence of SPA in the GenBank.A 876 bp SPA gene was amplified from the genomic DNA of Staphylococcus aureus(ATCC12598),which encodes 292 amino acids including five IgG-binding regions of E,D,A,B and C.A prokaryotic expression vector pET28a(+)-SPA was then constructed and recombinant protein with molecular weight of 35.0 ku was expressed successfully with IPTG induction.The expressed product was purified by IgG-affinity chromato-graphy.Western-blot analysis indicated that the purified SPA possessed biological activity. |
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| Keywords: | staphylococcal protein A gene cloning prokaryotic expression affinity chromatography |
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