中文摘要：

目的利用哺乳动物细胞表达HIV-1病毒样颗粒,为HIV-1病毒的生物学研究和进一步设计HIV-1中和优势表位预防性疫苗奠定基础。方法以293T细胞作为宿主细胞,利用磷酸钙沉淀法将带有HIV-1病毒被膜蛋白gp160基因的pcDNA3.1/BaL gp160真核表达载体和带有HIV-1核心蛋白（gag）的pVRC3900真核表达载体共转染至293T细胞中,收集培养上清,经蔗糖不连续梯度离心,收集病毒样颗粒,经磷钨酸负染,透射电镜观察结果。结果以磷酸钙沉淀DNA转移技术,EGFP真核表达质粒为靶基因,293T细胞为宿主细胞,最佳转染效率达30%以上;将pcDNA3.1/ BaL gp160（env）和pVRC3900（gag）真核表达质粒共转染,宿主细胞成功表达目的蛋白,转染细胞培养上清经蔗糖密度梯度离心,电镜观察可见自行装配的HIV病毒样颗粒。结论在真核细胞中表达HIV-1病毒样颗粒。

英文摘要：

Aim To express HIV-1 virus-like particle（VLP） in mammalian cells, which may facilitate the further research on HIV-1 biological characteristics and prophylactic vaccine aimed at neutralizing epitope on the basis of VLP. Methods pcDNA3.1/BaL gp160 with virus envelope gene gp160 and pVRC3900 with virus core gene gag were cotransfected into 293T cells using calcium phosphate precipitation. The supernatants of cultures were collected and centrifuged by noncontinuous sucrose centrifugation. The VLPs were detected under transmission electron microscope after negative staining by phosphotungstic acid. Results After cotransfection of pcDNA3.1/BaL gp160 with virus envelope gene gpl60 and pVRC3900 with virus core gene gag into 293T cells, interested proteins were successfully expressed. The expressed proteins could be auto-assembled into VLP, which were observed under electron microscope. Conclusion HIV-1 VLP has been successfully expressed and assembled in host cells, which can be greatly helpful in further research on neutralizing predominant epitope prophylactic vaccines based on VLP.