Construction of mammalian cell expression vector of human leukocyte antigen A and B gene from peripheral blood mononuclear cells and analysis of its sequence

Objective To construct mammalian cell expression vector of human leukocyte antigen A,B gene. Methods Total cell RNA was extracted from peripheral blood mononuclear cell and the cDNA was amplified by RT PCR; The cDNA fragments were directly inserted into Pbluescript KS(+/-) and the recombinant PBS HLA A,B were identified by restriction endonucleases digestion and sequencing; Finally the fragments of HLA A,B were subcloned into pcDNA3 with restriction enzymes Hin dⅢ and Bam HⅠ.Results The recombinant mammalian cell expression vectors of pcDNA3 HLA A,B were constructed, the sequences of the insert were identical to the published sequen ces encoding HLA A*3001 and HLA B*27052 gene. Conclusion The recombinant mammalian cell expression vectors of pcDNA3 HLA A*3001 and pcDNA HLA B*27052 were successfully constructed. [