玫瑰叶畸形病毒外壳蛋白基因原核表达与抗血清制备  

Prokaryotic expression and antiserum preparation of coat protein of rosa rugosa leaf distortion virus

作　　者：张秀琪 聂张尧 李梦林 苏江月 丁泽慧 张宗英 韩成贵 王颖 ZHANG Xiuqi;NIE Zhangyao;LI Menglin;SU Jiangyue;DING Zehui;ZHANG Zongying;HAN Chenggui;WANG Ying(Key Laboratory of Pest Monitoring and Green Management,Ministry of Agriculture and Rural Affairs,College of Plant Protection,China Agricultural University,Beijing 100193,China)

机构地区：[1]中国农业大学植物保护学院,农业农村部作物有害生物监测与绿色防控重点实验室,北京100193

出　　处：《植物保护》2024年第4期230-234,241,共6页Plant Protection

基　　金：国家级大学生创新训练项目(201910019028)。

摘　　要：玫瑰叶畸形病毒(rosa rugosa leaf distortion virus,RrLDV)属于番茄丛矮病毒科Tombusviridae天竺葵环斑病毒属Pelarspovirus,能够引起月季叶片畸形、黄化以及提前衰老等症状,是危害月季的病毒之一。为建立玫瑰叶畸形病毒的快速检测方法,采用热硼酸盐法提取发病月季叶片总RNA,通过RT-PCR扩增该病毒外壳蛋白基因,连接至原核表达载体pDB-His-MBP上,导入大肠杆菌Escherichia coli BL21以IPTG,诱导蛋白表达,获得浓度为8 mg/mL的重组蛋白,制备了兔多抗血清。Western blot检测血清效价为1∶2048000,当效价为1∶1000时,灵敏度为1∶1024。血清学检测结果与RT-PCR检测结果一致,表明该抗血清可用于RrLDV的检测,有利于检测该病毒的发生和分布。Rosa rugosa leaf distortion virus(RrLDV)belongs to the genus Pelarspovirus in the family Tombusviridae,which can infect roses and cause symptoms such as leaf deformity,yellowing and premature aging.In order to establish a rapid detection method for detecting RrLDV,total RNA of susceptible rose leaves was extracted by hot borate method.The gene which encodes coat protein of the virus was amplified by RT-PCR and constructed to the prokaryotic expression vector pDB-His-MBP.The vector was transformed into Escherichia coli BL21 and the expression of the recombinant protein was induced by IPTG.The recombinant protein with a concentration of 8 mg/mL was obtained and the polyclonal anti-rabbit antiserum was produced.Western blotting showed the titer of the antiserum was 1∶2048000 and its sensitivity was 1∶1024 when the antiserum was diluted to 1∶1000.The serological detection result was consistent with RT-PCR,indicating that the antiserum can be used to detect the occurrence and distribution of RrLDV.

关 键 词：玫瑰叶畸形病毒 外壳蛋白 原核表达 抗血清制备 

分 类 号：S436.81[农业科学—农业昆虫与害虫防治]