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利用基因工程技术分离出含有Ⅰ型单纯疱疹病毒（HSV－1）包装信号序列和HSV－1基因组复制起点序列的片段，表达外源基因所必需的启动子序列以及其下游作为报告基因的lacZ基因片段，构建成质粒型HSV－1载体pHSVL。用脂质体介导的方法可将该质粒转染入经辅助病毒HSV－1tsK株超感染的Vero细胞，并将其包装成HSV－1假型病毒颗粒，可如天然病毒一样再感染传代细胞和神经细胞并在其中进行基因表达。在pHSVL的基础上，我们还构建了一种可同时表达两种外源基因的质粒型疱疹病毒载体pHLCL，即在pHSVL中插入第二个转录单位：HCMVIE启动子和其下游的荧光素酶基因（lucgene），由此获得的pHLCL病毒接种传代细胞证明它可在其中同时表达两种报告基因．本研究成功地构建了两种质粒型HSV－1载体，其独特的基因转移方式使之在神经生理、病理及神经系统疾病的基因治疗的实验研究中都具有广阔的应用前景。
A gene fragment containing the HSV-1 packaging signal sequence and the HSV-1 genome origin of replication, the promoter sequence necessary for the expression of foreign genes and the lacZ gene downstream thereof as a reporter gene were isolated by genetic engineering Fragment, constructed into plasmid vector HSV-1 pHSVL. This plasmid can be transfected into Vero cells superinfected with the helper virus HSV-1 tsK strain by liposome-mediated methods and packaged into HSV-1 pseudotyped virus particles, which can be re-infected with passaged cells as the native virus And nerve cells and gene expression therein. On the basis of pHSVL, we also constructed a plasmid herpesvirus vector pHLCL that can express two foreign genes at the same time. That is, a second transcription unit was inserted into pHSVL: HCMVIE promoter and its downstream luciferase gene (Lucgene) and the resulting thus obtained pHLCL virus seeded cells showed that it can express both reporter genes simultaneously. In this study, two plasmid-based HSV-1 vectors were successfully constructed, and their unique gene transfer methods have broad application prospects in the gene therapy of neurophysiological, pathological and neurological diseases.