中国兽医科学2011，Vol.41Issue(10)：1034-1040,7.
肾型传染性支气管炎病毒的鉴定和结构蛋白的表达
Characterization of a nephropathogenic infectious bronchitis virus strain and expression of its structural proteins
摘要
Abstract
This research was conducted to study an isolate of infectious bronchitis virus（Ck/Jiangsu/DS10/2008,DS10） from layer poultry farm in Jiangsu province and to provide information for the control of the disease.DS10 virus isloate was characterized by a series of biological test.The S,M and E genes of DS10 virus were amplified by reverse transcription polymerase chain reaction（RT-PCR）.The amplified fragments were sequenced and phylogenic tree of the genes were constructed with genes of other viruses from GenBank.Subsequently,these genes were cloned into the pFastBac 1 vector of Bac-to-Bac baculovirus expression system and transfected into Sf9 cell line with the expression and biological activity detected using indirect immunofluorescentassay（IFA） and agar immunodiffusion,respectively.Phylogenetic analysis showed that there was no significantly correlation between different genes of DS10 virus.DS10 virus was identified as nephropathogenic strains by its biological properties.Indirect fluorescence assay（IFA） showed that S,M and E proteins were expressed in Sf9 cell line.The agar immunodiffusion assay showed that S and M proteins expressed in Sf9 cell had positive precipitation band with serum against IBV,while the E protein did not being found precipitation.Results indicated that the expressed S and M proteins had powerful biological activity.The low rate in virus total protein and small molecular weight of E protein may lead to invisible of precipitation band in neutralization with serum.For the vaccine strain of IBV at present can not provid full protection against to variant viruses challenge,the expressed proteins of S,M and E in the present study afford fundamental data for the research of its biological activity and development of diagnosis antigen and subunit vaccine.关键词
传染性支气管炎病毒/S基因/M基因/E基因/杆状病毒表达系统Key words
infectious bronchitis virus/S gene/M gene/E gene/Bac-to-Bac baculovirus expression system分类
农业科技引用本文复制引用
唐应华,吴培培,宫玉珍,王永伟,于漾,张道华,何家惠,侯继波..肾型传染性支气管炎病毒的鉴定和结构蛋白的表达[J].中国兽医科学,2011,41(10):1034-1040,7.基金项目
江苏省农业科技自主创新项目[cx10（450)] （）
江苏省自然科学基金面上项目（BK2009328) （）
