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时间:2020-03-25
《猪胞内劳森氏菌GXNN株4个抗原性候选基因抗原性分析.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、广西农业科学2010,41(1):62-65GuangxiAgriculturalSciences猪胞内劳森氏菌GXNN株4个抗原性候选基因抗原性分析肖爱欢1,谢丽华2,廖承球-,兰家暖-,李茂宁-,侯韶毅1,陆仕勇3,黄伟坚”(1广西大学动物科学技术学院。南宁530005;2梧州市动物疫病预防控制中心,广西捂州543002;3广西农垦国有金光农场,南宁530042)摘要:参考GenBank上已发表的胞内劳森氏菌相关蛋白序列,设计了4对引物,分别扩增了4个抗原性候选基因(3个外膜蛋白基因和1个表面脂蛋白基因),并将4个抗原性
2、候选基因构建好原核表达载体后进行原核表达、SDS—PAGE电泳和Westernblotting分析。SDS-PAGE电泳检测初步确定重组融合蛋白pET32a—L10902、pET32a—L11022能够进行表达,分别得到53、37kDa的条带;Westernblotting分析则初步确定pET32a—U1022具有抗原性。研究结果首次验证胞内劳森氏菌外膜蛋白基因是否具有抗原性,为诊断试剂盒及基因工程疫苗的研制提供理论基础。关键词:胞内劳森氏菌;抗原性;候选基因;SDS-PAGE电泳;Westernblotting;猪中图分类
3、号:$858.28文献标识码:A文章编号:1002—8161(2010)01--0062--04AnalysisofantigenicityinfourantigeniccandidategenesofLawsoniaintracellularisGXNNstrainXIAOAi-huanl,XIELi—hua2,LIAOCheng—qiul,LANJia-nuanl,LIMao-nin91,HOUShao-yil,LUShi—yongs,HUANGWei-jianl+(1CollegeofAnimalScienceandT
4、echnology,GuangxiUniversity,Nanning530005,China;2WuzhouPreventionandControlCenterofAnimalEpidemicDisease,Wuzhou,Gnangxi543002,China;3Gu哪,iJingnangStateFarm,Nanning530042,China)Abstraet:AccordingtotheassociatedproteinsequencesofLawson函intracellularispubhshedinC,enBa
5、nk。fourpairsofpfimemweredesignedandthegenesofthreeoutermembraneproteinsandoneectallipoproteinwereamplified.Afterconstructingtheprokaryoticexpressionvectorsforfourantigeniccandidategenes,theprokaryoticexpression,SDS-PAGEeleetrophoresisandWesternblottinganalysiswe弛pe
6、rformed.Theresultsshowedthattworecombinationfusionpro-reinspET32a-L10902andpET32a-L11022couldbeexpressedand57kDaand37kDabandWasobtainedbySDS-PAGEelectrophoresis,respectively.TheWesternblottinganalysisresultsshowedthatpET32a-L11022hadantigenieity.Thesere-suitsforthe
7、firsttimeprovedthatwhetherouternlembrsneproteinsofLawsoniaintraeellularishadantigenieityornot,andwouldprovidetheoreticalbasisfordevelopingdiagnostickitandgeneticallyengineeredvaccine.Keywords:Lawson/aintrace]lularis;antigenicity;candidategene;SDS-PAGEelectrophoresi
8、s;Westernblotting猪增生性肠炎(Porcineproliferativeentempathy。PPE)又称猪回肠炎(Porcineileitis),是由胞内劳森氏菌(Lawson/aIna'aceIlularis。LI)引起的一种接触性传染病。以回肠及结盲处近端的结肠、盲肠
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