[Construction of MHC I gene retroviral expression vector and its expression in hematopoietic cells].

ABSTRACT

OBJECTIVE: To construct the retroviral expression vector of BALB/c mouse H-2Dd gene and study its expression in C57BL/6 mouse hematopoietic cells (MHC). METHODS: A retroviral vector pMSCV encoding H-2Dd gene was transduced into the packaging cell line PT67 by lipofectin, and the hematopoietic cells of C57BL/6(H-2Dd negative) were infected by the above viral supernatant. Reverse transcriptase-polymerase chain reaction and flow cytometry were employed to examine the expression of H-2Dd on the infected cells. RESULTS: The cDNA encoding H-2Dd was correctly inserted into the vector pMSCV, as confirmed by restriction endonuclease digestion. The H-2Dd gene was integrated into the C57BL/6 mouse hematopoietic cell genome and expressed on the cell surface. CONCLUSION: Recombinant H-2Dd of BALB/c mouse retroviral expression vector has been successfully constructed and its expression obtained in C57BL/6 mouse hematopoietic cells, which may facilitate further study of the function of MHC in transplantation immunology.