Expression of HCV coreAnt Construction of recombinant lentivirus.docVIP

 PAGE \* MERGEFORMAT 16 Expression of HCV coreAnt Construction of recombinant lentivirus [Abstract] Objective To construct the expression of HCV core area of recombinant lentiviral, to explore the HCV infection of the virus core protein on liver stem cell transdifferentiation. Methods PCR system extends each other template primers T vector cloning to obtain HCV core Ant cloning, and ligated into the pLenti6/V5 D TOPO plasmid by pLenti6/V5 D HCV core Ant, together with the pLP / VSVG, pLP1 and pLP2 four plasmids were transfected 293ET cells was expressed HCV core Ant lentiviral . collection of viral supernatant, a lot of amplification, detected by immunohistochemistry gene in Hela cells. in a similar way the expression of EGFP was constructed and the recombinant lentiviral titer was measured. Results cloned HCV core Ant genes restriction analysis and sequencing confirmed that the results are correct; use of the expression of HCV core Ant Hela cells infected with recombinant lentivirus, immunohistochemical experiments confirmed the expression of HCV core cytoplasm there. the expression of EGFP using recombinant lentiviral infection of 3T3 cells to see the green fluorescence Construction lentiviral vector effective. Conclusions In vitro recombination by molecular cloning expression of HCV core was successfully prepared recombinant lentiviral for further study of hepatitis C virus mechanisms underlying cancer. [Keywords:] Hepatitis C virus core protein; recombinant lentivirus; molecular cloning ABSTRACT: Objective To construct a recombinant lentiviral vector for HCV core Ant and then study its effect on the transdifferentiation of hepatic stem cells. Methods The HCV core Ant was obtained by PCR of two primers template with each other and T vector cloning method, and then subcloned to pLenti6/V5 D TOPO. The restriction endonuclease and T4 DNA ligase were used to construct the vector. pLenti6/V5 D HCV core Ant and the ViraPowerTM PackagingMix (containing thr