Screening Of Highly Efficient Recombinant Transmembrane Peptide - Vp3 And Its Inhibitory Effect On Hela Cells

Objective: To construct the soluble expression PTD4-apoptin, R9-apoptin, hC-SOD3-apoptin fusion protein three prokaryotic expression vector, to detect the apoptosis of Hela cancer cells, and filter out the effect of suppressing the strongest, with high activity and strong cell-permeable penetrating peptide-apoptin.Methods: pGEX-6p-1 vector to construct pGEX-6p-1-PTD4-apoptin, pGEX-6p-1-R9-apoptin and pGEX-6p-1-hC-SOD3-apoptin prokaryotic expression vector, make three kinds of expression coli expression vectors E.coli BL21(DE3) in. Induced by IPTG, GST affinity column, after ultrafiltration was concentrated to give a high concentration of the fusion protein. Hela cells and incubated by MTT assay PTD4-apoptin, R9-apoptin and three hC-SOD3-apoptin fusion protein on apoptosis in Hela cells.Results: We successfully constructed PTD4-apoptin, soluble expression hC-SOD3-apoptin and R9-apoptin fusion protein in E. coli transformed by purified to obtain a high concentration of the fusion protein, the cancer cells were incubated together with Hela, PTD4-apoptin, hC-SOD3-apoptin and R9-apoptin three kinds of fusion proteins promote cancer cell apoptosis Hela. PTD4-apoptin inhibition rates of Hela 82.34%, hC-SOD3-apoptin inhibition rates of Hela 66.60%, R9-apoptin inhibition rates of Hela 45.03%.Conclusion: The expression of PTD4-apoptin, R9-apoptin, hC-SOD3-apoptin three kinds of soluble fusion protein, and to inhibit cancer cells Hela.