Hefferon K L, Khalilian H, Xu H, AbouHaidar M G
Department of Botany, University of Toronto, Ontario, Canada.
J Gen Virol. 1997 Nov;78 ( Pt 11):3051-9. doi: 10.1099/0022-1317-78-11-3051.
Transgenic potato plants were generated which express a dicistronic mRNA corresponding to the 8 kDa protein and coat protein (CP) genes of potato virus X (PVX). Northern blot analysis of RNA isolated from these plants indicated that both the 8 kDa protein and the CP are translated from this dicistronic mRNA. Expression of both of these proteins in transgenic plants was demonstrated by Western blot analysis, and suggests that translation of CP takes place either by initiation by internal entry of ribosomes or by a termination/reinitiation mechanism. CP was detected in protoplasts electroporated with RNA transcripts of the dicistronic construct in the presence or absence of a stable hairpin structure at the 5' terminus, suggesting that the former model is more likely. Similar results were obtained when a DNA fragment containing the PVX CP leader sequence was placed between two reporter genes in the transient assay system. These results suggest that potexvirus CP expression may take place from the genomic RNA as well as from the subgenomic RNA.
构建了转基因马铃薯植株,其表达对应于马铃薯X病毒(PVX)8 kDa蛋白和外壳蛋白(CP)基因的双顺反子mRNA。对从这些植株中分离的RNA进行Northern印迹分析表明,8 kDa蛋白和CP均从该双顺反子mRNA翻译而来。通过Western印迹分析证实了这两种蛋白在转基因植株中的表达,这表明CP的翻译可能通过核糖体内部进入起始,或者通过终止/重新起始机制进行。在用双顺反子构建体的RNA转录本进行电穿孔的原生质体中,无论5'末端是否存在稳定的发夹结构,均检测到了CP,这表明前一种模式更有可能。在瞬时分析系统中,当将含有PVX CP前导序列的DNA片段置于两个报告基因之间时,也获得了类似的结果。这些结果表明,马铃薯X病毒CP的表达可能发生在基因组RNA以及亚基因组RNA上。