Abstract：The present paper mainly studies the human CD154 protein the nucleus expression. Carries on the massive expressions under the GST nucleus expression most superior condition, and after the GST column chromatographic analysis purification, obtains the purity high goal protein to use in the immunology experiment. This research is directs the thing according to the goal protein CD154 gene sequence design synthesis specificity, PCR to expand increases the gene, and inserts to the fusion protein nucleus expresses in carrier pGEX-4T-1, obtains the reorganization to express material particle pGEX-4T-1/CD154,Reorganizes the material particle transformation backwoods E.coli DH5-α; cell with this, the transformed colony after BamHⅠ、EcoRⅠ;The double enzyme cuts the appraisal. And through uses the different IPTG induction density and the different raise temperature and the time, causes GST to fuse the protein DH5-α; to obtain the biggest expression at the backwoods E.coli, and gathers the sample to carry on the SDS-PAGE electrophoresis appraisal to express the product. Through to transforms after the backwoods coli to carry on the SDS-PAGE electrophoretic analysis, discovered the obvious appearance 54kD protein leucorrhea. Also best expression condition: 37℃, IPTG induction density 0.2mM, induction time 4h.This studied constructs the GST nucleus to express the material particle successfully, and expressed the GST fusion protein in the backwoods coli, might use in a deeper level after the depuration GST fusion protein the immunity experiment.
keywords： GST fusion protein, expression, E.Coli , SDS-PAGE
