Smith R H, Zhao Y, O'Callaghan D J
Department of Microbiology and Immunology, Louisiana State University Medical Center, Shreveport 71130-3932.
J Virol. 1993 Feb;67(2):1105-9. doi: 10.1128/JVI.67.2.1105-1109.1993.
We have previously reported that the equine herpesvirus 1 (EHV-1) XbaI G restriction fragment (nucleotides 1436 to 7943 relative to the left terminus of the EHV-1 genome [Kentucky A strain]) is required in combination with the EHV-1 immediate-early (IE) gene to achieve significant activation of two representative EHV-1 late promoter-chloramphenicol acetyltransferase (CAT) recombinants in transient expression assays. In this report, we demonstrate that the XbaI G-encoded UL3 gene (an ICP27 homolog) provides a trans-acting factor which acts (in combination with the EHV-1 IE gene product) to increase reporter gene expression directed by an EHV-1 late promoter-CAT recombinant plasmid. We show that cloned copies of UL3 can successfully substitute for the XbaI G fragment in CAT assays and that stop codon insertion within the UL3 open reading frame inhibits the ability of UL3 to activate reporter gene expression in trans.
我们先前曾报道,在瞬时表达试验中,马疱疹病毒1型(EHV-1)XbaI G限制性片段(相对于EHV-1基因组[肯塔基A株]左末端的核苷酸1436至7943)需要与EHV-1立即早期(IE)基因结合,才能显著激活两个具有代表性的EHV-1晚期启动子-氯霉素乙酰转移酶(CAT)重组体。在本报告中,我们证明XbaI G编码的UL3基因(一种ICP27同源物)提供了一种反式作用因子,该因子(与EHV-1 IE基因产物结合)作用以增加由EHV-1晚期启动子-CAT重组质粒指导的报告基因表达。我们表明,UL3的克隆拷贝可以在CAT试验中成功替代XbaI G片段,并且在UL3开放阅读框内插入终止密码子会抑制UL3反式激活报告基因表达的能力。