人类疱疹病毒7 U41的部分功能鉴定与分析

Identification of the function of human herpesvirus 7 U41

目的 :对人类疱疹病毒 7(humanherpesvirus 7,HHV 7)U4 1的部分功能进行分析鉴定 ,为药物治疗学探索提供依据。方法 :免疫荧光检测U4 1蛋白的细胞内定位 ,RT PCR检测U4 1RNA的表达时相 ,DNA结合试验检测U4 1蛋白对单、双链DNA的不同结合能力。结果 :免疫荧光检测显示 ,无论在HHV 7感染细胞或HHV 7U4 1转染细胞 ,U4 1蛋白都是仅在其胞核中弥漫性的分布。在分别加入环己酰亚胺和磷甲酸作为蛋白合成和核酸合成抑制剂感染细胞中 ,U4 1的表达时相检测结果显示U4 1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U4 1蛋白能结合单链DNA而不能结合双链DNA。结论 :U4 1的主要功能是保持DNA模板的合适构型 。

AIM: To identify the function ORF (open reading frame) U41 of human herpesvirus 7(HHV 7). METHODS: Immunofluorescent detection was used to show the localization of the U41 protein in cells. The expression kinetics of the U41 gene in the presence of cycloheximide (CHX) and phosphomoformic acid (PFA), which are inhibitors of protein synthesis and of viral DNA synthesis respectively, was detected with RT PCR. DNA binding assay was used to detect the single and double DNA strands binding ability of U41 protein. RESULTS: The U41 protein was expressed as an early protein in the HHV 7 infected cells and localized only in the nuclei of infected cells. The U41 protein was able to bind to a single strand DNA cellulose column but not to double strand DNA cellulose column from DNA binding assay. CONCLUSION: The major function of U41 may be to keep the DNA template in the optimal conformation for DNA elongation and for viral DNA synthesis.