Scheets K, Khosravi-Far R, Nutter R C
Department of Plant Pathology, Oklahoma State University, Stillwater 74078.
Virology. 1993 Apr;193(2):1006-9. doi: 10.1006/viro.1993.1216.
A full-length cDNA clone (pMCM41) was constructed to contain the exact 5' end of MCMV behind a T7 RNA polymerase promoter and a Smal site at the 3' end. Uncapped RNA synthesized from pMCM41 has the exact 3' end of viral RNA (vRNA) but is missing the cap found on vRNA. This RNA was infectious in protoplasts from black Mexican sweet (BMS) maize (Zea mays) suspension cultures. Uncapped transcripts were also infectious when inoculated onto maize plants and produced an infection indistinguishable from vRNA-inoculated plants. Capped pMCM41 transcripts which initiated at position +2 of the cDNA clone, as well as capped or uncapped RNA synthesized from a clone containing an extra G between the T7 promoter and the 5' end of MCMV sequence (pMCM721), were less infectious than uncapped pMCM41 transcripts in BMS protoplasts. The transcripts one nucleotide longer or shorter than uncapped pMCM41 transcripts were not able to infect maize plants.
构建了一个全长cDNA克隆(pMCM41),使其在T7 RNA聚合酶启动子后包含MCMV的精确5'端,在3'端有一个Smal位点。从pMCM41合成的无帽RNA具有病毒RNA(vRNA)的精确3'端,但缺少vRNA上的帽结构。这种RNA在来自墨西哥黑甜(BMS)玉米(玉米)悬浮培养物的原生质体中具有感染性。当接种到玉米植株上时,无帽转录本也具有感染性,并产生与接种vRNA的植株无法区分的感染。在BMS原生质体中,从cDNA克隆的+2位置起始的加帽pMCM41转录本,以及从在T7启动子和MCMV序列5'端之间含有一个额外G的克隆(pMCM721)合成的加帽或无帽RNA,其感染性低于无帽pMCM41转录本。比无帽pMCM41转录本长一个核苷酸或短一个核苷酸的转录本无法感染玉米植株。