大蒜A病毒CP基因的原核表达及抗血清制备.pdf

生物技术通报 -研究报告· BIoTEcHNoLoGYBULLETlN 2011年第7期 大蒜A病毒CP基因的原核表达及抗血清制备 杨宇1 韦传宝1’2 华俊雅1 刘春云1 吴琪瑶3 摘 要： 均为98％。将Gary-ACP基因插入表达载体pSBET，在大肠杆茵BL2l(DE3)Plys 和5％一20％SDS．PAGE两次纯化，免疫小鼠获得抗CP血清，Westernblotting分析表明确定制备的抗体时CP具有高度特异 性，ELISA检测表明制备的抗体能够与天然病毒离子结合，因此可以作为该病毒的检测。 关键词： 大蒜A病毒CP基因原核表达抗血清制备 A Prokaryotic ofGarlicvirusCPGeneand Expression ItsAntiserum of Preparation YulWeiChuanba01·2Hua Liu Wu Yang Junyal ChunyunlQiya03 andPharmaceutical Anhui 237012； (1Biological EngineeringCollege，WestUniverdty，Liuan 2AnhaiProvincial BiomimerticSensorand Laboratoryof DetectingTechnology，luuan237012； 3／#e＆如眦College，AnhuiUnioerslty，日咖230039) wfl8 to GarlicvirusALiuanisolate．The showsthatGarV·Ashared Abstract： designed multiplealigment Specificprimer amplify 98％一99％nucleotideacidsidentitiesand98％aminoacidsidentitieswiththetwo ofCP onGenBank．Then reported sequencesgenes theCP wa8insertedinto and inEscherichiacoli Estrain．The was gene pSBETexpressed BL21(DE3)plysobjectproteinpurifiedby theCPWasraisedinmouseandits 12％SDSand SDS．Theantiserum fireflysubsequently5％一20％gradient against issuitablefor detection。 wDJ5confirmedWestern resultindicatedthattheantiserum GarV·A specificity by blottinganal