| 副猪嗜血杆菌OMP P2基因的克隆与表达 |
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| 引用本文： | 李鹏,李军星,曹珺,张国龙,李玉峰,宋艳华,王先炜,姜平.副猪嗜血杆菌OMP P2基因的克隆与表达[J].中国兽医学报,2012,32(2):212-215. |
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| 作者姓名： | 李鹏 李军星 曹珺 张国龙 李玉峰 宋艳华 王先炜 姜平 |
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| 作者单位： | 1. 南京农业大学农业部动物疫病诊断与免疫重点开放实验室,江苏南京210095／长江大学动物科学学院,湖北荆州434025
2. 南京农业大学农业部动物疫病诊断与免疫重点开放实验室,江苏南京,210095 |
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| 基金项目： | 浙江省科技农业项目(2007C22045);江苏省教育厅产业发展基金资助项目(JH09-1);农业部公益行为专项基金资助项目(200903036-04) |
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| 摘 要： | 利用已分离的菌株HB070412,根据NCBI上GenBank中的HPS序列（ZP_02477753）设计了1对引物,用PCR方法扩增了副猪嗜血杆菌外膜蛋白中的穿孔素前体蛋白P2基因（OMP P2）,并克隆到载体pMD18-T（T-Vec-tor）,序列测定结果表明OMP P2基因全长1 077bp,序列同源性分析表明该基因相当保守,与所报道的HPS OMPP2基因之间的核苷酸序列同源性为98.1%,氨基酸序列同源性为95.5%。用pGEX-6p-1构建了原核表达载体pGEX-P2,在大肠杆菌中进行了高效表达,表达蛋白约占菌体总蛋白的50%,主要为包涵体形式。SDS-PAGE电泳结果显示表达的融合蛋白约为65 000,Western blot结果表明该表达蛋白具有抗原性。
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| 关 键 词： | 副猪嗜血杆菌 OMP P2基因 克隆 原核表达 |
Cloning and expression of the OMP P2 gene of Haemophilus Parasuis |
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| LI Peng,LI Jun-xing,CAO Jun,ZHANG Guo-long,LI Yu-feng,SONG Yan-hua,WANG Xian-wei,JIANG Ping.Cloning and expression of the OMP P2 gene of Haemophilus Parasuis[J].Chinese Journal of Veterinary Science,2012,32(2):212-215. |
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| Authors: | LI Peng LI Jun-xing CAO Jun ZHANG Guo-long LI Yu-feng SONG Yan-hua WANG Xian-wei JIANG Ping |
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| Institution: | 1*(1.Key Laboratory of Animal Diseases Diagnostic and Immunology,Ministry of Agriculture,College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;2.College of Animal science,Yangtze University,Jingzhou,Hubei 434025,China) |
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| Abstract: | A pair of specific primers was designed based on the published sequences of the OMP P2 gene of Haemophilus Parasuis(HPS),the OMP P2 gene was amplified from the isolated strain HB070412 of HPS by PCR.The PCR amplified DNA fragment was cloned into pMD18-T.The sequencing results indicated that this gene was 1 077 bp and quite conservative.The result of sequence analysis of OMP P2 gene of HB070412 indicated that the homology among other HPS strains would be from 95.5% to 98.1%.The OMP P2 gene was ligated into pGEX-6p-1 to get the expressing vector of pGEX-P2 which was then transformed into E.coli BL21.The result of SDS-PAGE shows the expressed protein is about 65 000,and the recombinant protein accounts for 50% of total cell protein and in inclusion body form.Western blot results indicate that this protein possesses biological activity. |
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| Keywords: | Haemophilus Parasuis OMP P2 gene cloning prokaryotic expression |
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