马疱疹病毒1型调控蛋白对SV40早期启动子的调控  

Role of equine herpesvirus 1 regulatory proteins in regulating SV40 early promoter

作　　者：高俊[1,2] 刘荻萩[2] 马月[2] 王晓钧[1,2] 

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]中国农业科学院哈尔滨兽医研究所兽医生物国家重点实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2016年第4期298-302,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金青年项目(31402202)

摘　　要：为研究马疱疹病毒1型(EHV-1)6种调控蛋白(极早期调控蛋白IEP、早期调控蛋白EICP0、EICP22、EICP27、IR2P和晚期调控蛋白ETIF)对猴空泡病毒40(SV40)早期启动子(SV40EIp)的调控作用,本研究将这6种EHV-1调控蛋白真核表达重组质粒分别与SV40EIp萤火虫荧光素酶重组报告质粒共转染RK13细胞,并同时转染作为内控的海肾荧光素酶报告质粒,通过检测两种独立的报告酶与对应底物反应后的发光强度,得到SV40EIp活性相对值。从而评价EHV-1 6种调控蛋白对SV40EIp活性影响。结果表明:ETIF对SV40EIp活性影响不显著;IR2P对SV40EIp活性具有下调作用;IEP、EICP0、EICP22、EICP27对SV40EIp活性均具有上调作用;而对于含增强子的SV40EIp(SV40EIp E),这4种调控蛋白也能够上调SV40EIp E活性,其中EICP22上调作用最强。而且,EICP22和EICP27能够协同上调SV40EIp的活性。此外,EICP22与EHV-1早期基因启动子转录因子TBP、EICP27与TBP及EICP22与EICP27的免疫共沉淀试验(Co-IP)结果表明:仅EICP27可以与TBP发生相互作用,提示EICP22并非与EICP27或者TBP直接结合而实现协同上调作用,其协同上调作用还存在其他的作用方式。To evaluate the effect of equine herpesvirus 1（EHV-1） regulatory proteins including immediate early regulatory protein（IEP）, early regulatory proteins（EICP0, EICP22, EICP27, IR2P） and late tegument protein（ETIF） on the trans-activated activity of simian virus 40 early promoter（SV40EIp）, the six recombinaant expressing plasmids of the EHV-1 regulatory proteins were co-transfected with the p GL3-SV40（expression of luciferase under control of SV40EIp） in RK13 cells, respectively, and subjected to the dual luciferase（DLR） assays to measure the activity of the luminescent reaction with Renilla luciferase as control.The results showed that ETIF had no significant impact on the activity of SV40 EIp, and the SV40 EIp was even down regulated by IR2 P. In contrast, IEP, EICP0, EICP22 and EICP27 were able to trans-activate SV40 EIp. In addition, the SV40 EIp E（SV40EIp,contained an enhancer sequence in the promoter region）, was able to be enhanced by EICP22, IEP, EICP27 and EICP0. Moreover,EICP22 and EICP27 had the ability of synergistic enhancing SV40 EIp. Further identification showed that EICP27 interacted with TATA binding protein（TBP）, but not EICP22, indicating that the synergistic enhancing mechanism for SV40 EIp by EICP22 and EICP27 was not directly interacted between the two regulatory proteins, which need to be further investigation.

关 键 词：EHV-1 EICP22 EICP27 调控作用 SV40早期启动子 

分 类 号：S852.65[农业科学—基础兽医学]