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番茄环斑病毒5’RNA1保守片段的克隆及病毒检测

5'RNA1 CONSERVATIVE SEGMENT CLONING OF TOMATO RINGSPOT NEPOVIRUS AND ITS DETECTION
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摘要 提取感染番茄环斑病毒TomatoRingspotNepovirus(TomRSV)的番杏 (TetragoniaExpansa)和葡萄品种赤霞珠(Vitis.cv.CabernetSauvignon)嫩叶及其阴性对照的总RNA ,根据NCBI中报道的TomRSV病毒四个分离物RNA15’端保守片段设计引物 ,对提取的总RNA进行RT -PCR扩增 ,获得长 4 5 0bp特异性片段 ,并克隆到PMD18-T载体中构建PT -TomRSV重组载体。以PT -TomRSV载体为阳性对照 ,以无毒赤霞珠叶片为阴性对照 ,对带毒赤霞珠葡萄叶片作番茄环斑病毒巢式PCR检测。 Total RNA was extracted from the leaves of Tetragonia Expansa inoculated by Tomato ringspot nepovirus(TomRSV) ,one-year-old Carbernate Sauvignon plants infected by TomRSV and the leaves of T.Expansa and Carbernate Sauvignon uninfected by TomRSV. Using the primers designed according to the 5'RNA1 conservative segment of the four isolates of TomRSV, the 450bp segment was got by RT-PCR amplifying the extracted RNA, then cloned it into PMD18-T vector to construct the recombinant vector PT-TomRSV. With the positive control of vector PT-TomRSV and negative control of Carbernate Sauvignon leaves uninfected TomRSV, the Carbernate Sauvignon leaves infected TomRSV were detected by Nest-PCR.
出处 《山东农业大学学报(自然科学版)》 CSCD 北大核心 2003年第3期361-365,共5页 Journal of Shandong Agricultural University:Natural Science Edition
关键词 番茄 环斑病毒 5’RNA1保守片段 克隆 病毒检测 Tomato ringspot nepovirus nest-PCR clone detection
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  • 1成卓敏,病毒学报,1992年,8卷,80页
  • 2胡伟贞,植物检疫,1991年,5卷,249页

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