Abstract: To construct the siRNAs expressing plasmid aiming at the H-2K^d gene and to explore the H-2K^d expression in mouse LAK cells and function of H-2K^d with the RNA interference technology. MethodsFirst, a target sequence of the H-2K^d gene was selected, then according to the sequence, two complementary oligonucleotides were lisated into psilencer 3.0-H1, which was transformed into DH5a bacteria, and then it was amplified and purified to get the plasmid. The purified plasmid was identified by gel electrophoresis and sequencing. H-2K^d-targeting siRNA plasmid was used in spleen lymphocytes of BALB/C mice. Flow cytometry(FCM) was used to determine the expression of H-2K^d on the transfected and control cells. ResultsGel electrophoresis and sequencing results showed that the plasmid was about 2.8kb which was identical with the positive control, and the sequence was identical with what had been inserted. Also siRNA treatment significantly inhibited the expression of the targeted protein. Conclusion siRNAs expression plasmids aiming at the H-2K^d gene have been successfully constructed and could inhibit the H-2Kd expression.

Key words: Plasmid, H-2K^dgene, siRNA