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目的 通过比较小鼠与人重排活化基因 2 (RAG2 )启动子 ,试图寻找与小鼠RAG2启动子特异性结合的转录因子。方法 采用表达luciferase的报告基因载体检测启动子的活性。采用EMSA(electrophoresismobilityshiftassay)检测与启动子结合的转录因子。结果 小鼠RAG2启动子 - 6 0 - 4 1区域存在富G的GA盒子 ,而人RAG2启动子在相应位置却是富A区。突变实验结果显示 ,GA盒子是小鼠RAG2启动子完整活性所必须的。EMSA结果显示 ,Sp1 Sp3结合在小鼠RAG2启动子 - 6 0 - 4 1区域 ,并且Sp1能够协同Pax 5、c Myb活化小鼠RAG2启动子。结论 尽管小鼠与人RAG2启动子同源性很高 ,但它们结合的转录因子和功能有所不同。
Objective To search for transcription factors that specifically bind to the mouse RAG2 promoter by comparing the mouse and human rearrangement activating gene 2 (RAG2) promoters. Methods The reporter gene vector expressing luciferase was used to detect the promoter activity. EMSA (electrophoresismobilityshiftassay) detection of promoter binding transcription factor. As a result, a G-rich G box was present in the mouse RAG2 promoter-6 0-4 1 region, whereas the human RAG2 promoter was a region rich in A at the corresponding position. The results of the mutation experiment show that GA box is necessary for the complete activity of mouse RAG2 promoter. EMSA results show that Sp1 Sp3 binds to the mouse RAG2 promoter -60-4 region, and Sp1 is capable of activating the mouse RAG2 promoter in cooperation with Pax 5, c Myb. Conclusion Although mouse has high homology with human RAG2 promoter, the binding factors and functions of mouse RAG2 are different.