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猪繁殖和呼吸综合症病毒河南地方株ORF5/6基因的克隆及真核表达

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摘要 应用RT-PCR方法分别扩增出猪繁殖与呼吸综合征病毒(PRRSV)河南地方株的ORF5和0RF6基因。将扩增基因克隆入真核表达载体pcDNA3.0中,设计不同引物自行构建重组质粒PcDNA-ORF5、ORF6和pcDNA-ORF5/6。运用磷酸钙法将重组质粒分别瞬时转染293T细胞,经流式细胞和westbloting试验分析表明共表达重组质粒PcD-NA-ORF5/6表达蛋白能够与PRRSV的阳性血清发生特异性反应,而pcDNA-ORF5-ORF6在293T细胞很难检测到蛋白表达。该结果表明蛋白质生物活性和其蛋白质空间构象有一定关系,也为下一步进行构建PRRSV假型病毒来研究ORF5/6蛋白的结构与功能的关系奠定了基础。
作者 党占国 李志勇 夏平安 陈溥言 崔保安 李伟娟 王中明
机构地区 河南农业大学牧医工程学院 焦作市动物疫病预防控制中心 南京农业大学动物医学院
出处 《畜牧兽医科技信息》 2008年第8期22-25,共4页 Chinese Journal of Animal Husbandry and Veterinary Medicine
基金 中国博士后基金项目资助(No20060390944)
关键词 PRRSV ORF5/6基因 克隆 真核表达
分类号 S858.28 [农业科学—临床兽医学]
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参考文献15

  • 1Dea S,et al. Current knowledge on the structural proteins of porcine reproductive and respiratory syndrome (PRRS) virus: comparison of the North American and European isolates[J].Arch Virol. 2000,145(4):659-688.
  • 2Meier WA,et al. Gradual development of the interferon,gamma response of swine to porcine reproductive and respiratory syn dronle virus infection or vaccination[J].Virology.2003,309(1):18-31.
  • 3Mengeling WL,et al. Safety and efficacy of vaccination of pregnant gilts against pocine reproductive and respiratory syndrome[J].J Vet Res, 1999,60(7):796-801.
  • 4Bother A, et al.Heterologous challenge with porcine reproductive and respiratory syndrome(PRRS) vaccine virus:no evidence of rectivation of previous European-type PRRS virus infectious[J]. Vet Microbiol,1999,68(3-4): 187-195.
  • 5Nielsen TL, et al. Examination of virus shedding in semen from vaccinated and from previously infected boars after experimental challenge with porcine reproductive and respiratory syndrome virus[J].Vet Microbiol.1997,54(2):101-102.
  • 6Nilubol D, et al. The effect of a killed porcine reproductive and respiratory syndrome virus (PRRSV) vaccine treatment on virus shedding in previously PRRSV infected pigs[J].Vet Mierobiol.2004 19,102(1-2):11-18.
  • 7Yoon K J, et al. Field isolate of porcine reproductive and resp iratorysyndrome virus (PRRSV) vary in their susceptibility to antibody dependent enhancement (ADE) of infects[J]. Vet Micrbiol.1997. 55:277 -287.
  • 8Meulenberg J J , et al. Characterization of p roteins encoded by ORFs 2 to 7 of Lelystad virus[J]. Virology, 1995,206: 155-163.
  • 9KapurV, et al. Genetic Variation in porcine reproductive and respiratory syndrome virus isolates intheMidwesteru United States[J].J Virol,1996,77: 1271-1276.
  • 10萨姆布鲁克J 弗里其EF 曼尼阿蒂斯T.分子克隆实验指南[M]第2版[M].北京:科学出版社,1996.880-98.

二级参考文献19

  • 1谢京国,方六荣,肖少波,姚林,潘永飞,陈焕春.双基因共表达的“自杀性”DNA疫苗载体的改造及其体外表达特性[J].中国兽医学报,2006,26(6):583-585. 被引量：2
  • 2Wensvoort G,Terpstra C,Pol J M.Mystery swine disease in the Netherlands:the isolation of Lelystad virus.Veterinary Quarterly,1991,13:121-130.
  • 3Dea S,Gagnon C A,Mardassi H.Current knowledge on the structural proteins of porcine reproductive and respiratory syndrome (PRRS) virus:comparison of the North American and European isolates.Archive of Virology,2000,145:659-688.
  • 4Leitner W W,Ying H,Restifo N P.DNA and RNA-based vaccines:principles,progress and prospects.Vaccine,2000,18:765-775.
  • 5Tubulekas I,Berglund P,Fleeton M,Liljestrom P.Alphavirus expression vectors and their use as recombinant vaccines:a minireview.Gene,1997,190:191-195.
  • 6Liljeqvist S,Stahl S.Production of recombinant subunit vaccines:protein immunogens,live delivery systems and nucleic acid vaccines.Journal of Biotechnology,1999,73:11-33.
  • 7Fleeton M N,Liljestrom P,Sheahan B J,Atkins G J.Recombinant Semliki Forest virus particles expressing louping ill virus antigens induce a better protective response than plasmid-based DNA vaccines or and inactivated whole particle vaccine.Journal of General Virology,2000,81:749-758.
  • 8Herweijer H,Wolff J A.Self-amplifying vectors for gene delivery.Advanced Drug Delivery Reviews,1997,27:5-16.
  • 9Hariharan M J,Driver D A,Townsend K,Brumm D,Polo J M,Belli B A,Catton D J,Hsu D,Mittelstaedt D,McCornmack J E,Karavodin L,Dubensky T W,Chang S M,Banks T A.DNA immunization against herpes simplex virus:enhanced efficacy using a Sindbis virus-based vector.Journal of Virology,1998,72:950-958.
  • 10Meulenberg J J,Hulst M M,De Meifer E J,Moonen P L,Den Bestern A,De Kluyver E P,Wensvoort G,Moormann R J.Lelystad virus,the causative agent of porcine epidemic abortion and respiratory syndrome (PEDRS),is related to LDV and EAV.Virology,1993,192:62-72.

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畜牧兽医科技信息
2008年 第8期

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