An Experimental Study On The Gene Synthesization,Prokaryotic Expression Plasmid Construction And Prokaryotic Expression Of Improved Fusion Protein TAT-PTD-VP3

VP3 is a protein produced by chicken anemia virus(CAV) which can induce apoptosis in malignant tumor cells. Because the VP3 is too big to pass the cell membrane automatically, researchers have to express the protein in the tumor cells through transport the gene of VP3 on an eukaryotic expression plasmid into the tumor cells. However, the operation to transport gene into tumor cells is difficult, the efficiency of eukaryotic expression is too low to get a satisfied treat result, and the risk to transport a therapeutic gene into the body of patients, all of these limit the clinical using of this protein, so if we can produce the VP3 in the lab, then transport it into tumor cells through some effective ways, those problem of eukaryotic expression would be resolved. TAT-PTD is a protein with low molecular weight which can transport another protein, the molecular weight of which is from 15 X 10~3～2 X 10~5, into cells with high efficiency, Improved TAT-PTD is a transporter of protein devised according to the TAT-PTD, the efficiency of which to transport protein into cells is 30 times higher than that of TAT-PTD. So if composed VP3 with improved TAT-PTD, the problem of how to transport VP3 into tumor cells will be resolved.Our research include synthesizing the gene of fusion protein TAT-PTD-VP3, composing the prokaryotic expression plasmid pGEX-6p-1-TAT-PTD/ VP3 and...