vpr基因在GFP荧光基因转染SHIV病毒模型中的作用

【目的】通过在vpr基因区不同部位插入EGFP基因,探讨vpr非结构基因的改变对SHIV病毒复制及感染能力的影响。【方法】利用分子克隆的方法,将EGFP基因插入SHIV基因组中的调节基因vpr基因中,最终获得相应的SHIV XJDC6431-EGFP全长克隆,然后在细胞水平检测该病毒感染活性及荧光蛋白表达情况。【结果】通过定点突变及在vpr基因不同部位插入GFP报告基因,检测改造后的质粒虽表达绿色荧光,但经细胞水平检测后均来得到具有感染活性的病毒颗粒。【结论】vpr基因定点突变可使全长SHIV病毒复制大大降低,共至失去感染活性。

Effect of vpr gene in GFP fluorescence gene transfection SHIV virus model

【Objective】Through the insertion of the expressing green fluorescent protein（EGFP） into different parts of vpr gene region,to observe the impact of change of changes in non-structural genes vpr on simian-human immunodeficiency virus（SHIV） replication and infection ability.【Methods】 By using the method of molecular cloning,to insert the EGFP gene into regulating gene vpr of the genome of SHIV,then to obtain the corresponding SHIV XJDC6431- EGFP full-length cloning,and finally to detect the virus infection activity and fluorescence protein expression at a cellular level.【Results】 Through site-directed mutagenesis and the insertion of GFP into different parts of vpr gene region,green fluorescence appeared in modified plasmid,but active virus particle with infection was not detected at the cellular level.【Conclusions】 Vpr gene-directed mutagenesis could greatly reduce the replication of total length SHIV virus,and even make it lose infection activity.