Optimization Oflentivirus Vector Systemand Its Application In Gene Over-Expression And RNAinterference

As a way of transfection,lentivirus vector can introduce exogenous gene into host cells,and do no harm to host cell genome function.In addition,the ablity to infect no-dividing cells makes lentivirus a favourite tool in scientific and medical research.Recently,lentivirus vetor system has been used in the research of gene functions viaover-expression and RNAi.To clinical trials,a great improvement has been made in the treatment of tumor by RNAi with the aid of lentivirus vector system.In consideration of the advantages of lentivirus vector system,our main work is to optimize the lentivirus infection system for the follow-up study.During this work,several basic conditions of virus packaging were optimized,such as plasmid and reagents ratio,the number of transfection cell,pretreating and expression time.Simultaneously,in order to make the lentivirus system an effective one,GFP was taken asreport protein to optimize cell density,Moi,infection,and expression time.The optimized lentivirus vetor system was used in the study of function of the T6SS effector EvpP in J774A.1 cells during the co-cultivation of Edwardsiella piscicidav and J774A.1 cells,and EvpP was found to act as an inhibitor in the activation of inflammasome.Moreover,with the help of lenvirus vector system,it was proven that adoral plays an important role in the targeting of AMP to CW-2 and MDA-MB-468 using RNA interference(RNAi).