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旨在构建HSV-1HF株的扩增子载体,研究其在不同血清型HSV辅助下的包装通用性。经酶切HF株的BAC-HSV-1,获得oriS和pac元件并测序。以pSilencer2.0-U6为骨架,以DsRed为报告基因构建HSV-1HF株的扩增子载体,利用脂质体2000转染扩增子载体至Vero细胞,分别应用HSV-1HF株和HSV-2HG52辅助HSV-1扩增子载体进行包装,待产生细胞病变效应后取上清,再次感染Vero细胞,观察Vero细胞内红色荧光蛋白表达情况。本研究首次构建了HSV-1HF株的扩增子载体,鉴定了HSV-1HF株oriS和pac元件,HSV-1HF株扩增子载体可以被HSV-1HF株和HSV-2HG52株包装并扩增。
Aim To construct the amplicon carrier of HSV-1HF strain and study its packaging versatility with the assistance of HSV of different serotypes. The oriS and pac elements were obtained and sequenced by restriction enzyme digestion of BAC-HSV-1. Using pSilencer2.0-U6 as the backbone and DsRed as the reporter gene to construct the HSV-1HF amplicon vector, the amplicon was transfected into Vero cells by lipofectamine 2000. The HSV-1HF strain and HSV-2HG52 Auxiliary HSV-1 amplicon carrier packaging, to be cytopathic effect after the supernatant was taken, re-infected with Vero cells observed Vero cells red fluorescent protein expression. In this study, we constructed the amplicon vector of HSV-1HF strain for the first time and identified the oriS and pac elements of HSV-1HF strain. The HSV-1HF amplicon vector can be packaged and amplified by HSV-1HF strain and HSV-2HG52 strain.