Construction And Expression Of Recombinant Adenovirus Carrying HIV-1 Vpr Gene And Preliminary Study On Its Biological Functions

Objective:To study on the biological effect of Viral protein R on the biological behaviour of C8166 cell, providing important insights into Vpr function and its mechanism. And a foundation for further research on the proteomics floating of CD4~+ T cell induced by Vpr.Methods:Recombinant adenovirus Ad-vpr was constructed. Vpr gene was linked to pAdTrack-CMV plasmid and then homologous recombination was performed in E. coli BJ5183 between pAdTrack-CMV- vpr and adenovirus backbone plasmid pAdEasy. The obtaining recombinant vector was transfected into 293 cells and packaged out recombinant adenovirus Ad-vpr. C8166 cell infected by Ad-vpr was observed by fluorescene microscopy. Identification of Vpr expression in C8166 cell was confirmed with western blotting. C8166 cell membrane integrity change, cell cycle arrest and apoptosis that induced by Vpr were ditected by flow cytometry. Electronic microscope was used to observe morphologic change of C8166 cell infected by Vpr.Results:The results of identification showed the insertion was right and the recombinant adenovirus contained the vpr gene. Vpr expression in Ad-vpr infected C8166 cell was...