中文摘要：

目的：研究单纯疱疹病毒Ⅱ型（HSV-2）潜伏相关转录体（LAT）开放读码框1（ORF1）的表达特点及其对Vero细胞活性的影响。方法：双酶切和测序验证本实验室构建的HSV-2 LAT ORF1真核表达载体pEGFP-ORF1,并以转染试剂盒Xfect介导其转染至Vero细胞,通过RT-PCR和绿色荧光蛋白检验其在细胞中的表达,用MTT法进行细胞活性分析。结果：重组质粒表达的融合蛋白主要集中细胞核,而空质粒表达的绿色荧光蛋白在细胞核和细胞质中分布均匀;重组质粒对Vero细胞没有损伤作用。结论：HSV-2 LAT ORF1影响了绿色荧光蛋白的分布,可降低空质粒对细胞的损伤作用;其作用位点可能主要定位在细胞核中,为阐明HSV-2 LAT ORF1在潜伏复发中的功能奠定了实验基础。

英文摘要：

Objective： To study the expression and characteristics of HSV-2 LAT ORF1 and its effects on Vero cells activities in vitro.Methods： The laboratory constructed HSV-2 LAT ORF1 eukaryotic expression vector pEGFP-ORF1 was verified by double en-zymes digestion and sequencing.Then the recombinant plasmid was transfected into Vero cells mediated by the kit Xfect.The expression of the recombination plasmid in the cells was observed by fluorescence microscopy and the expression of ORF1 was verified by RT-PCR.The cells activities were analyzed by MTT.Results： Fusion protein expressed by the recombinant plasmid is mainly exists in the nucleus,while the empty plasmid expressing green fluorescent protein distributed evenly in the nucleus and cytoplasm.The recombinant plasmid had no harmful to the Vero cells.Conclusion： HSV-2 LAT ORF1 can reduce the damage caused by empty plasmid and the target may be mainly exists in the nucleus.The function of ORF1 in Vero cells may has lay the basis for further research on the mechanism of HSV-2 LAT latency and reactivation.