摘要
应用免疫荧光检测U41蛋白的细胞内定位,RT-PCR检测U41RNA的表达时相,DNA结合试验检测U41蛋白对单、双链DNA的不同结合能力,对人类疱疹病毒7(humanherpesvirus7,HHV-7)U41的部分功能进行分析鉴定。免疫荧光检测显示,无论在HHV-7感染细胞或HHV-7U41转染细胞,U41蛋白都是仅在其胞核中呈弥漫性分布。在分别加入环己酰亚胺(cycloheximide)和磷甲酸(phosphonoformicacid)作为蛋白合成和核酸合成抑制剂的感染细胞中,U41的表达时相检测结果显示,U41蛋白在HHV-7感染细胞中是一早期表达蛋白。通过DNA结合试验发现,U41蛋白能结合单链DNA而不能结合双链DNA。提示U41的主要功能是保持DNA模板的合适构型,以便DNA的延伸和病毒DNA的合成。
Immunofluorescent detection was used to show the localization of the U41 protein in cellsThe expression kinetics of the U41 gene in the presence of cycloheximide(CHX)and phosphonoformic acid(PFA),which are inhibitors of protein synthesis and of viral DNA synthesis respectively,was detected with RTPCRDNA binding assay was done to detect the single and double DNA strands binding ability of U41 proteinThe results showed that the U41 protein was expressed as an early protein in the HHV-7 infected cells and localized only in the nuclei of the infected cellsUsing DNA binding assay,we found that the U41 protein was able to bind to a singlestrand DNA cellulose column but not to doublestrand DNA cellulose columnThese findings strongly suggested that the major function of U41 was to keep the DNA template in the optimal conformation for DNA elongation and be required for viral DNA synthesis
出处
《病毒学报》
CAS
CSCD
北大核心
2002年第4期303-306,共4页
Chinese Journal of Virology
