胡葱黄条病毒外壳蛋白的原核表达与抗血清的制备  

Prokaryotic expression and antiserum preparation of coat protein of Shallot yellow stripe virus

作　　者：罗朝鹏[1] 林林[1] 陈炯[2] 吴云锋[1] 陈剑平[2] 

机构地区：[1]西北农林科技大学植保学院,陕西杨凌712100 [2]浙江省农业科学院病毒学与生物技术研究所,浙江杭州310021

出　　处：《浙江农业学报》2007年第3期147-150,共4页Acta Agriculturae Zhejiangensis

基　　金：973前期专项(2006CB708209);国家自然科学基金项目(30200008);浙江省重大招标项目(2003C12023)

摘　　要：设计特异性引物PCR扩增得到胡葱黄条病毒(SYSV-O)的全长外壳蛋白(CP)基因,构建原核表达载体pET32a-SYSV-O CP,转化大肠杆菌诱导表达。结果表明,SYSV-O CP基因在大肠杆菌中得到了高效表达,分子量约为46 kDa。用纯化表达产物免疫小鼠制备抗血清,Western blot检测结果表明,抗血清与SYSV-O的CP发生了强烈的特异性反应。使用该抗血清进行了田间样品的带毒检测。Specific primers were designed and used in amplification of coat protein（CP） gene of shallot yellow stripe virus from onion （SYSV-O） in Shaanxi Province. PCP product was cloned into the pE332a vector, a 46 kDa protein was expressed. This protein was purified and used to immune mouse for antiserum preparation. Western blot analysis confirmed that the antiserum reacted strongly and specifically to CP of SYSV. This antiserum was also used to detect SYSV infections in the samples from shaanxi province.

关 键 词：胡葱黄条病毒 原核表达 抗血清 

分 类 号：Q344.13[生物学—遗传学]