Todd J W, Passarelli A L, Miller L K
Department of Genetics, University of Georgia, Athens 30602-2603.
J Virol. 1995 Feb;69(2):968-74. doi: 10.1128/JVI.69.2.968-974.1995.
We report the identification of four additional genes of the Autographa californica nuclear polyhedrosis virus involved in expression from a late baculovirus promoter in transient expression assays. Three of these genes, p35, 39K, and p47, have been previously described. The role of the p35 gene product in late gene expression may be related to its ability to block apoptosis, since two other baculovirus genes also known to block apoptosis, Cp-iap and Op-iap, were able to functionally replace p35 in the transient expression assay. The requirement for p47 in this assay confirms its role in late gene expression, a role previously established by characterization of a temperature-sensitive mutant of p47, while the requirement for 39K may be related to its known association with the virogenic stroma. The fourth gene identified as a late expression factor gene, lef-11, was located immediately upstream of 39K and is predicted to encode a 13-kDa polypeptide. When plasmids containing these 4 genes were cotransfected with plasmids containing the 14 genes previously identified as late gene expression factors, the level of expression from the late capsid promoter was similar to that observed for a library of clones representing the entire viral genome. The genes provided by these 18 plasmids thus represent the viral genes necessary and sufficient to support expression from a late viral promoter in this transient expression assay.
我们报告了在瞬时表达试验中鉴定出的另外四个与苜蓿银纹夜蛾核型多角体病毒(Autographa californica nuclear polyhedrosis virus)晚期杆状病毒启动子表达相关的基因。其中三个基因,即p35、39K和p47,此前已有描述。p35基因产物在晚期基因表达中的作用可能与其阻断细胞凋亡的能力有关,因为另外两个已知能阻断细胞凋亡的杆状病毒基因Cp-iap和Op-iap,在瞬时表达试验中能够功能性替代p35。在该试验中对p47的需求证实了其在晚期基因表达中的作用,这一作用先前已通过对p47温度敏感突变体的表征得以确立,而对39K的需求可能与其已知与病毒发生基质的关联有关。被鉴定为晚期表达因子基因的第四个基因lef-11,位于39K的紧邻上游,预计编码一个13 kDa的多肽。当含有这4个基因的质粒与含有先前鉴定为晚期基因表达因子的14个基因的质粒共转染时,晚期衣壳启动子的表达水平与代表整个病毒基因组的克隆文库所观察到的水平相似。因此,这18个质粒提供的基因代表了在该瞬时表达试验中支持晚期病毒启动子表达所必需且充分的病毒基因。