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[Objective]The effect of baicalin on Staphylococcus aureus thermostable nuclease enzyme nuc gene and its activity was analyzed.[Methods]The minimum inhibitory concentration of baicalin against Staphylococcus aureus was determined with the double dilution method,the impact of the drug on the activity of thermostable nuclease enzyme was determined with the method of Toluidine Blue DNAse Agar and the effect of baicalin on the expression of Staphylococcus aureus nuclease resistant bacteria nuc gene was detected with RT-PCR.[Results]The minimum inhibitory concentration of baicalin against Staphylococcus aureus was 0.156 mg/mL.The activity of Staphylococcus aureus thermostable nuclease enzyme could be completely inhibited the baicalin solution at the containing of 40.000-5.000 mg/mL and there was obvious inhibition to the activity of Staphylococcus aureus thermostable nuclease enzyme while the concentration of baicalin was diluted to 0.078mg/mL,which was significant difference with that of CK(P<0.01).The specific band of 867bp was successfully amplified with synthetic primers and its sequencing result was completely identical with the sequence of standard strain mu50.While the concentration of baicalin solution was 2.500 mg/mL the expression of Staphylococcus aureus thermostable nuclease enzyme nuc gene could be reduced.[Conclusions]The baicalin might inhibit the expression of Staphylococcus aureus thermostable nuclease enzyme nuc gene and reduce the activity of its heat-resisting.