Shippam C, Wu X, Stewart S, Theilmann D A
Department of Plant Science, University of British Columbia, Vancouver, Canada.
Virology. 1997 Jan 20;227(2):447-59. doi: 10.1006/viro.1996.8334.
opep-2 is an Orgyia pseudotsugata multicapsid nucleopolyhedrovirus (OpMNPV) early gene in the ie1-ie2 gene region for which there is no homolog in either the archetype virus, Autographa californica MNPV, or Bombyx mori NPV. opep-2 is transcribed immediately upon infection as three mRNAs which initiate from a early gene motif (TATA-N27-CAGT). The expression of multiple transcripts at very early times postinfection has only been previously described for the baculovirus early gene ie1, which produces spliced mRNAs. However, distinct from ie1, the multiple mRNAs of opep-2 are due to multiple termination sites and not splicing. Western blot analysis of steady-state levels of OPEP-2 showed that in OpMNPV-infected Ld652Y cells maximum levels are obtained at 8-12 hr postinfection (p.i.) prior to DNA replication. By 48 hr p.i. OPEP-2 is shut off and is undetectable. To aid in elucidating the function of this OpMNPV-specific gene an opep-2 deletion mutant was generated and was compared to wild-type virus to determine if its absence affects viral growth in Ld652Y tissue culture cells.
opep - 2是一种黄杉毒蛾多核衣壳核型多角体病毒(OpMNPV)的早期基因,位于ie1 - ie2基因区域,在原型病毒苜蓿银纹夜蛾多角体病毒(Autographa californica MNPV)或家蚕核型多角体病毒(Bombyx mori NPV)中均无同源物。opep - 2在感染后立即转录为三种mRNA,它们从一个早期基因基序(TATA - N27 - CAGT)起始。感染后极早期多个转录本的表达此前仅在杆状病毒早期基因ie1中被描述过,ie1产生剪接后的mRNA。然而,与ie1不同的是,opep - 2的多个mRNA是由于多个终止位点而非剪接产生的。对OPEP - 2稳态水平的蛋白质免疫印迹分析表明,在OpMNPV感染的Ld652Y细胞中,在感染后8 - 12小时(p.i.)DNA复制之前达到最高水平。到感染后48小时,OPEP - 2被关闭且无法检测到。为了有助于阐明这个OpMNPV特异性基因的功能,构建了一个opep - 2缺失突变体,并将其与野生型病毒进行比较,以确定其缺失是否会影响Ld652Y组织培养细胞中的病毒生长。