Prüfer D, Wipf-Scheibel C, Richards K, Guilley H, Lecoq H, Jonard G
Institut de Biologie Moléculaire des Plantes du CNRS, Strasbourg, France.
Virology. 1995 Dec 1;214(1):150-8. doi: 10.1006/viro.1995.9945.
A full-length cDNA of cucurbit aphid-borne yellows virus (CABYV) has been constructed and expressed either as an in vitro transcript, under control of a bacteriophage T7 RNA polymerase promoter, or in vivo, under control of the cauliflower mosaic virus 35S promoter in an agroinfection vector. The biological activity of the cloned cDNA was demonstrated by the ability of its in vitro transcript to replicate in protoplasts and of the agroinfection vector to infect agroinoculated plants. Virus in the agroinfected plants cold be transmitted by the aphid vectors Myzus persicae and Aphis gossypii. The specificity of luteovirus RNA packaging was investigated by replacing (1) the CABYV coat protein gene (and the overlapping ORF5) by the corresponding region of potato leafroll luteovirus or (2) the CABYV readthrough domain by the readthrough domain of beet western yellows luteovirus. The resulting chimeric transcripts replicated in protoplasts and produced virions.
已构建了葫芦科蚜传黄化病毒（CABYV）的全长cDNA，并将其作为体外转录本进行表达，该转录本受噬菌体T7 RNA聚合酶启动子控制，或者在体内表达，受农杆菌感染载体中花椰菜花叶病毒35S启动子控制。克隆的cDNA的生物活性通过其体外转录本在原生质体中复制的能力以及农杆菌感染载体感染农杆菌接种植物的能力得以证明。农杆菌感染的植物中的病毒可由蚜虫介体桃蚜和棉蚜传播。通过用马铃薯卷叶病毒的相应区域替换（1）CABYV外壳蛋白基因（和重叠的ORF5）或（2）用甜菜西方黄化病毒的通读结构域替换CABYV通读结构域，研究了黄症病毒RNA包装的特异性。所得的嵌合转录本在原生质体中复制并产生病毒粒子。
