Use Of T7 Phage Display Technique Experimental Study Of Hepatitis B Virus The Pres1 Interaction Protein

PARTâ… EXPRESSION PURIFICATION AND CHARACTERIZATION OF FUSION HEPAITIS B VIRUS SURFACE ANTIGEN PRES1 EXPRESSED IN ESCHERCHIA COLIObjective: To construct the prokaryotic expression vector with HBV PreS1 genes, express it highly in E.coli to get purified PreS1 fusion protein and then identify the protein's characterization to further establish a foundation for selecting PreS1-interacted proteins from cDNA expression library of human hepatic cells.Methods: PCR was deployed to amplify PreS1 gene segment containing Hindâ…¢and Xbaâ… endoenzyme sites. After double enzyme digestion, prokaryotic expression vectors pGST-MOLUC and PCR products were conjuncted by T4DNA ligase and converted into E.coli JM109 to construct and biopan the recombinant plasmids which were transformated into Eco-BL21(DE3) later. After IPGT induction, the fusion proteins were dealt with SDS-PAGE electrophoresis analysis and Western-blot detection. Then use glutathione-Sepharose 4B gelatum as filler to further purify the fusion proteins with affinity chromatograph.