| PCV-2陕西株ORF2基因的克隆、分析及原核表达 |
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| 引用本文： | 邢福珊,许信刚,童德文,王志昇,刘文静,宁蓬勃,张彦明.PCV-2陕西株ORF2基因的克隆、分析及原核表达[J].西北农林科技大学学报(社会科学版),2010(3):1-6. |
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| 作者姓名： | 邢福珊 许信刚 童德文 王志昇 刘文静 宁蓬勃 张彦明 |
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| 作者单位： | 西北农林科技大学动物医学院; |
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| 基金项目： | 陕西省科技攻关项目(2009K02-01); 教育部新世纪优秀人才支持计划(NCET-07-0701) |
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| 摘 要： | 【目的】克隆猪圆环病毒2型陕西分离株(PCV-2SX株)ORF2基因,并进行序列分析及原核表达。【方法】根据GenBank公布的PCV-2ORF2基因的核苷酸序列,设计并合成1对特异性引物,应用PCR方法扩增PCV-2SX株ORF2全长基因,将其克隆入pGEM-T载体中,进行测序及序列分析。然后,将ORF2基因亚克隆入原核表达载体pET-32a中,在大肠杆菌BL21中进行IPTG诱导表达,对表达产物进行SDS-PAGE和Western-blotting鉴定。【结果】扩增到了702bp的PCV-2ORF2全长基因。序列分析结果表明,PCV-2SX株ORF2基因与广西分离株(EF675237,ChinaGX)和巴西分离株(DQ861802,am21)的核苷酸序列同源性均达98.0%,氨基酸序列同源性均达98.3%,与其他毒株的核苷酸序列和氨基酸序列同源性分别在89.9%～97.9%和88.5%～98.0%。SDS-PAGE可检测到分子质量约为48ku的融合蛋白,主要以可溶性蛋白形式存在。Western-blotting分析表明,重组蛋白可被PCV-2阳性血清所识别。【结论】成功克隆了PCV-2SX株ORF2基因,并进行了原核表达。
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| 关 键 词： | 猪圆环病毒2型 陕西分离株 ORF2基因 序列分析 原核表达 |
Cloning,sequence analysis and prokaryotic expression of ORF2 gene of PCV-2 Shaanxi strain |
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| XING Fu-shan,XU Xin-gang,TONG De-wen,WANG Zhi-sheng,LIU Wen-jing,NING Peng-bo,ZHANG Yan-ming.Cloning,sequence analysis and prokaryotic expression of ORF2 gene of PCV-2 Shaanxi strain[J].Journal of Northwest Sci-Tech Univ of Agr and,2010(3):1-6. |
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| Authors: | XING Fu-shan XU Xin-gang TONG De-wen WANG Zhi-sheng LIU Wen-jing NING Peng-bo ZHANG Yan-ming |
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| Institution: | XING Fu-shan,XU Xin-gang,TONG De-wen,WANG Zhi-sheng,LIU Wen-jing,NING Peng-bo,ZHANG Yan-ming(College of Veterinary Medicine,Northwest A&F University,Yangling,Shaanxi 712100,China) |
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| Abstract: | Objective] PCV-2 SX strain ORF2 gene was cloned,analyzed and expressed in E.coli.Method] According to the published complete nucleotide sequence of PCV-2 in GenBank,a pair of primers specific to the full-length of ORF2 gene were designed.The ORF2 gene fragment amplified by PCR was cloned into pGEM-T vector.The recombinant plasmid was sequenced and ORF2 gene was compared with other Genebank's PCV-2 strains.ORF2 gene segment was amplified from recombinant plasmid by PCR,then the ORF2 gene was cloned into ve... |
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| Keywords: | PCV-2 Shaanxi strain ORF2 gene sequence analysis prokaryotic expression |
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