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


CONCLUSIONS
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.
Wu
Lan-Xiao
LX
Department of Hematology, Zhujiang Hospital, First Military Medical University, Guangzhou 510282, China. wulx75@yahoo.com
Guo
Kun-Yuan
KY
Li
Jiang-Qi
JQ
Li
Yu-Hua
YH
Li
Ming
M
D000818:Animals; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005805:Genes, MHC Class I; D005822:Genetic Vectors / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D012190:Retroviridae / Q000235:genetics