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时间:2020-05-20
《水稻黑条矮缩病病毒外壳蛋白基因S10的原核表达、多克隆抗体制备及应用.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、中国水稻科学(ChinJRiceSei),2010,24(1):25~3Ohttp://www.ricesei.cnDO1:10.3969/j.issn.1001—7216.2010.O1.0525水稻黑条矮缩病病毒外壳蛋白基因S/O的原核表达、多克隆抗体制备及应用欧阳元龙1,2,3吴建祥2,熊如意3周益军3周雪平(南京师范大学生命科学学院,江苏南京210046;浙江大学农业与生物技术学院生物技术研究所,浙江杭州310029;。江苏省农业科学院植物保护研究所,江苏南京210014;’通讯联系人。E—mail:wujx@zju.edu.cn)Pro
2、karyoticExpressionofCoatProteinGeneSJ0ofRiceBlack—StreakedDwarfVirus,andPreparationandApplicationofItsPolyclonalAntibodyOUYANGYuan~longt·。,WuJian~xiang,,XIONGRu—yi。,ZHOUYi—jun。,ZHOUXue—ping。(CollegeofLifeScience,NanjingNormalUniversity,Nanjing210046,China;InstituteofBiotechno
3、logy,CollegeofAgriculture&Biotechnology,ZhejiangUniversity,Hangzhou3】0029,China;。InstituteofPlantProtection,JiangsuAcademyofAgriculturalSciences—Nanjing210014,China;Correspondingauthor。E-mail:tc,ujx@“edu.cn)0UYANGYuanlong,WUJianxiang,XIONGRu—yi。eta1.Prokaryoticexpressionofcoa
4、tproteingeneS10ofriceblack—streakeddwarfvirus,preparationandapplicationofitspolyclonalantibody.ChinJRiceSci,2010,24(1):25—30.Abstract:Theful1lengthcDNAofriceblack—streakeddwarfvirus(RBSDV)segmentl0(0)whichencodedcoatproteinwasclonedfromthevirusinfeetedricesamplesbyRT_PCR,ands
5、ubclonedintoaprokaryoticexpressionvectorpET32a.Therecombinantprokaryotieexpressionvector(pET-32a—CP)wasusedtotransformEscher&iacoliBI2l(DE3).A76kDTrKAfusionproteinwasobtainedwithinductionofIPTGandpurificationofNi+NTAaffinitycolumn.Thepurifiedrecombinantproteinwasusedtoimmuniz
6、erabbitsforproductionofpolyclonalantibodiesagainstthecoatproteinofRBSDV.Usingpoly—clonalantibodies.immunocaptureRT—PCRandDot—blotELISAwereestablishedforreliable,sensitiveandspecificdetectionofRBSDV.Thetwodetectionmethodsutilizedpolyclonalantibodiesprovidetechnicalsupportforth
7、ediagnosisofRBSDVdis—ease.Keywords:riceblack—streakeddwarfvirus;prokaryoticexpression;polyclonalantibody;immunoeaptureRT—PCR;dot—blotE1ISA欧阳元龙,吴建祥,熊如意,等.水稻黑条矮缩病病毒外壳蛋白基因0的原核表达、多克隆抗体制备及应用.中国水稻科学,2OlO,24(1):25—3O.摘要:用RT—PCR方法从感染水稻黑条矮缩病毒(riceblack-streakeddwarfvirus,RBSDV)水稻中克隆该病
8、毒的外壳蛋白基因S/0,然后将此外壳蛋白基因再豫克隆到原核表达载体PET一32a中构建成重组原核表达载体pET32a—CP。将重组表达载
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