Gene clone,recombination and expression of CTLA-4,TCRV?8 of human T lymphocyte / 西安交通大学学报(医学版)

ABSTRACT

Objective To clone CTLA-4 and TCRV?8 gene from T lymphocyte of thyroid of Graves' disease (GD) patients,recombine to form CTLA-4-TCRV?8 fusion gene and express the fusion protein.Methods CTLA-4 and TCRV?8 gene was cloned from T lymphocyte of thyroid of GD patients by RT-PCR.Then it was recombined with expression plasmid in order.The correct plasmids were obtained after the restriction analysis and DNA sequencing.Prokaryotic expression of the fusion protein in E.coli,SDS-PAGE and Western Blotting were used to verify the fusion protein.Results Restriction analysis and DNA sequencing confirmed the correct sequence and insertion site of the recombinant plasmid.The recombinant fusion protein was successfully expressed in E.coli,which was consistent with the predicted putative calculating molecular weight.Conclusion CTLA-4-TCRV?8 gene was constructed and expressed successfully,providing gene product and application theory for immune tolerance therapy of GD.