Characteirzation Of Gene Expression Regulated By CD4Gene Promoter And Enhancer Of Porcine

CD4is one of transmembrane glycoprotein, which is expressed on the thesurface of T helper cells.can be combined with MHCII molecules peptide area.Cloning and characterization of the CD4gene sequences of human,monkey,rat,rabbit, cattle,cats,dogs and other animals. The study of CD4geneexpression is controlled by a variety of regulatory sequences, including promoters,enhancers and silencer regulatory elements. Therefore, studying of CD4geneexpression characteristics and understanding the mechanism of various regulatoryelements of CD4gene has become a research focus.Study heterogeneous genepromoter to regulate the efficiency of exogenous gene express ion in human tissuetumor cells also has been a hot research, this study can provide the basis forestablishment of animal models of human tumor.Methods:The pig CD4gene promoter was focasted by bioinformatics means;then using the cloned CD4genen promoter, the lentivirus package was performed andthe lentiviral vector was constructed. Affter the packaged virus was obtained the titerof virus was detected and the293T cells and Jurkat cells were infected to performgreen fluorescent protein detection.The expression efficiencies of exogenous genes ofthe pigs CD4gene promoter and EF-1α promoter were compared. Composed of ninedifferent combinations of fluorescent protein expression vector with porcine, human,mouse CD4gene promoter and enhancer green,Transiently transfected293T, MCF-7,SW1116, HepG2and Hela cells.Efficiency comparison of three different species ofCD4gene promoter and enhancer in the regulation of gene expression in human cells,observe and relative green fluorescent protein expression.Results:The pig CD4gene promoter was successfully forecasted and thelentiviral vector was successfully constructed and the virus was packaged out. Thedection of titer of virus and infection of293T cells and Jurkat cells showed that thestart effect of GFP of two promoters in the cells were similar. Efficiency comparisonof three different species of CD4gene promoter and enhancer in the regulation of gene expression in human cells. prove the porcine CD4gene promoter and enhancerefficiency was significantly higher than the regulation of the mouse and human CD4gene promoter and enhancer efficiency in regulation of gene expression in humancells;Found no difference in the efficiency of porcine CD4gene promoter andenhancer regulate gene expression of the two CMV and EF-1α promoter.Conclusion: In this study, cloned porcine CD4gene promoter and enhancerregulates exogenous gene expression in human cells, More efficient than theregulation of expression of the mouse and human CD4gene promoter and enhancerregulation efficiency; The efficiency of porcine CD4gene promoter and enhancerregulate gene expression and regulation efficiency of the two CMV and EF-1αpromoter was no difference;the cloned pigs CD4gene promoter and enhancer doesnot have a population-specific.