【摘要】 为确认转录因子NF-κB对牛MMP9启动子的影响,利用P65过表达载体转染牛乳腺上皮细胞,检测其在蛋白水平对MMP9表达的影响;然后构建牛MMP9启动子双荧光素酶报告载体,转染Hela细胞后用双荧光素酶实验检测启动子活性,确定MMP9基因核心启动子区域,同时添加NF-κB抑制剂(Celastrol)确认P65对启动子活性的影响。结果为P65过表达能显著提高牛乳腺上皮细胞中的p-P65、P65和MMP9的蛋白表达;牛MMP9基因启动子上转录因子NF-κB结合位点在-420～-80 bp区域;NF-κB抑制剂显著降低牛MMP9基因启动子活性。因此转录因子NF-κB可直接靶向牛MMP9基因启动子区域并促进MMP9基因表达。更多还原

【Abstract】 In order to confirm the effect of transcription factor NF-κB on the bovine MMP9 promoter, the P65 overexpression vector was transfected to bovine mammary epithelial cells to detect the protein level of MMP9; then bovine MMP9 promoter dual luciferase reporter vectors were constructed, and the promoter activities were determined by dual luciferase assay after transfection of Hela cells. The core promoter region of MMP9 was determined too. NF-κB inhibitor(Celastrol) was added to test the effect of P65 on the promoter activity. The results showed that P65 overexpression can significantly increase the protein level of p-P65, P65 and MMP9 in bovine mammary epithelial cells; the binding site of NF-κB on the bovine MMP9 promoter was in the-420--80 bp region; NF-κB inhibitor significantly reduced the activity of bovine MMP9 promoter. Conclusion: NF-κB can directly target the promoter region of bovine MMP9 and promote the expression of MMP9.更多还原