MacFarlane S A, Brown D J
Virology Department, Scottish Crop Research Institute, Invergowrie, Dundee, UK.
J Gen Virol. 1995 May;76 ( Pt 5):1299-304. doi: 10.1099/0022-1317-76-5-1299.
A cDNA clone containing the complete coding region of RNA2 of the newly described TPA56 isolate of pea early-browning virus (English serotype, PEBV-E) has been obtained. A plasmid (pFLA56) containing this clone together with the 5' and 3' non-coding regions of PEBV isolate SP5 (the type isolate of PEBV-E) and flanked by the CaMV 35S promoter and NOS terminator is infectious when co-inoculated onto plants with pCaN1, an infectious clone of PEBV SP5 RNA1. Virus containing RNA2 derived from the cDNA clone of the TPA56 isolate is transmitted by Trichodorus primitivus nematodes, whereas virus containing RNA2 from a clone of the SP5 isolate is not transmitted. Sequencing revealed 11 differences out of 3374 nucleotides between the transmissible TPA56 and non-transmissible SP5 clones. However, only three of the base changes affected the amino acid sequences of virus gene products. A single, conservative amino acid change was present in the virus coat protein. Two non-conservative changes occurred in the protein of molecular mass 29.6 kDa expressed from an open reading frame located down-stream of the coat protein gene, suggesting that it has a function in nematode transmission and that changes in this protein prevent vector transmission of the SP5 isolate of PEBV.
已获得一个包含新描述的豌豆早褐病毒TPA56分离株(英文血清型,PEBV-E)RNA2完整编码区的cDNA克隆。一个质粒(pFLA56),包含该克隆以及PEBV分离株SP5(PEBV-E的典型分离株)的5'和3'非编码区,并由CaMV 35S启动子和NOS终止子侧翼包围,当与PEBV SP5 RNA1的感染性克隆pCaN1共同接种到植物上时具有感染性。含有源自TPA56分离株cDNA克隆的RNA2的病毒可由原始毛刺线虫传播,而含有来自SP5分离株克隆的RNA2的病毒则不能传播。测序显示,可传播的TPA56克隆和不可传播的SP5克隆在3374个核苷酸中有11个差异。然而,只有三个碱基变化影响病毒基因产物的氨基酸序列。病毒外壳蛋白中存在一个单一的保守氨基酸变化。在外壳蛋白基因下游的一个开放阅读框表达的分子量为29.6 kDa的蛋白质中发生了两个非保守变化,这表明它在线虫传播中具有功能,并且该蛋白质的变化阻止了PEBV SP5分离株的载体传播。