| 猪戊型肝炎病毒广西株ORF2三段连续基因编码蛋白的表达及抗原性分析 |
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| 引用本文： | 肖爱欢,梁保忠,黄伟坚,赵武,梁家幸,何颖,李茂宁,秦毅斌,李斌,侯韶毅,苏乾莲. 猪戊型肝炎病毒广西株ORF2三段连续基因编码蛋白的表达及抗原性分析[J]. 中国兽医科学, 2010, 40(6) |
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| 作者姓名： | 肖爱欢 梁保忠 黄伟坚 赵武 梁家幸 何颖 李茂宁 秦毅斌 李斌 侯韶毅 苏乾莲 |
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| 作者单位： | 肖爱欢,黄伟坚,李茂宁,侯韶毅(广西大学,动物科学技术学院预防兽医学广西区重点实验室,广西,南宁,530005;广西兽医研究所病毒研究室,广西,南宁,530001);梁保忠,赵武,梁家幸,何颖,秦毅斌,李斌,苏乾莲(广西兽医研究所病毒研究室,广西,南宁,530001) |
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| 基金项目： | 广西基本科研业务费专项 |
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| 摘 要： | 为了确定猪戊型肝炎病毒(HEV)ORF2抗原性片段的分布,参考中国T1株序列设计套式引物,从新鲜猪粪中扩增出HEV部分基因片段,长1725bp,位于ORF2第205～1929bp之间,命名为HEVORF2-A。根据HEVORF2-A序列设计了3对引物,分别扩增HEVORF2-A三段连续的基因片段,命名为HEVORF2-B1、HEVORF2-B2、HEVORF2-B3,分别位于HEVORF2-A第1～576bp,577～1149bp,1150～1725bp之间。将这4段扩增的基因片段分别与表达载体pET32a(+)连接,并转入BL21(DE3)pLysS感受态细胞,经IPTG诱导,SDS-PAGE分析。结果显示,经1mmol/LIPTG37℃诱导6h后,除pET32a-A在82.6ku处有微量融合蛋白表达外,pET32a-B1、pET32a-B2、pET32a-B3分别在41.3ku、41.5ku、41.8ku处有高水平的融合蛋白表达。用自然感染猪血清和北京万泰HEVELISA诊断试剂盒阳性对照血清进行Western-blot分析,结果3个短的融合蛋白均能检测到明显的条带,表明构建的3个表达载体具有良好的抗原性。
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| 关 键 词： | 猪戊型肝炎病毒 表达 抗原性 |
Expression and antigenic analysis of three recombinant continuous ORF2 proteins of swine hepatitis E virus Guangxi strain |
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| XIAO Ai-huan,,ZHAO Wu,LIANG Jia-xing,HE Ying,LI Mao-ning,QIN Yi-bin,LI Bin,HOU Shao-yi,SU Qian-lian,LIANG Bao-zhong,HUANG Wei-jian. Expression and antigenic analysis of three recombinant continuous ORF2 proteins of swine hepatitis E virus Guangxi strain[J]. Chinese Veterinary Science, 2010, 40(6) |
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| Authors: | XIAO Ai-huan ZHAO Wu LIANG Jia-xing HE Ying LI Mao-ning QIN Yi-bin LI Bin HOU Shao-yi SU Qian-lian LIANG Bao-zhong HUANG Wei-jian |
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| Affiliation: | XIAO Ai-huan1,2,ZHAO Wu2,LIANG Jia-xing2,HE Ying2,LI Mao-ning1,QIN Yi-bin2,LI Bin2,HOU Shao-yi1,SU Qian-lian2,LIANG Bao-zhong2,HUANG Wei-jian1(1.Key Laboratory of Preventive Veterinary Medicine of Guangxi Province/College of Animal Science and Technology,Guangxi University,Nanning 530005,China,2.Department of Virology,Guangxi Veterinary Research Institute,Nanning 530001,China) |
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| Abstract: | In order to determine the distribution of hepatitis E virus(HEV) ORF2 antigenic fragments,two pairs of primers were designed according to the sequence of China T1 strain and a fragment of ORF2 gene was amplified from fresh porcine feces.The fragment named as HEVORF2 A was 1 725 bp in size,located between nucleotide 205 and 1 929 of ORF2.Subsequently,three pairs of primers were designed based on HEVORF2 A,and three continuous gene fragments were amplified from HEVORF2-A,which were named as HEVORF2-B1,HEVORF2... |
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| Keywords: | ORF2 |
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