Objective:To construct the recombinant adenovirus carrying a short interfering RNA（siRNA）targeting MK（mouse killer） gene and identify its inhibitive effect in mouse uterine stromal cells.Methods:The siRNA sequence targeting MK gene was synthesized and cloned into the shuttle plasmid pSES-HUS to generate the vector pSES-HUS-MKsiRNA

which was later homogenously recombinanted with the adenovirus backbone plasmid pAdeasy-1 in Ecoli BJ5183.Then the recombinant adenovirus was transfected into HEK293 cells to be packaged and amplified.The primarily cultured mouse stromal cells were infected with this purified adenovirus and the expression levels of MK mRNA and protein were detected by RT-PCR and Western Blot.Results:The recombinant adenoviral vector containing MK siRNA was correctly constructed with an average titer of 6.8×10

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pfu/mL.Both the MK mRNA and protein levels were signifcantly decreased in the uterine stromal cells after 48 hours of infection with MK siRNA recombinant adenovirus.Conclusion:The generated recombinant adenovirus Adeasy-MK-siRNA was demonstrated to significantly abrogate the expression of MK at mRNA and protein level in mouse uterine stromal cells.