摘要
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 identify transcriptional factor specific for murine RAG2 (recombination activating gene 2) promoter by comparison of murine and human RAG2 promoter. Methods Luciferase reporter assay was used to detect the activity of promoter. EMSA was used to identify transcriptional factor. Results GA box exist in the -60/-41 region of murine RAG2 promoter, while rich A nucleotide region exist in human RAG2 promoter. Mutant assay result showed that GA box was necessary for the full activity of murine RAG2 promoter. EMSA result showed that Sp1/Sp3 bound to -60/-41 region of murine RAG2 promoter. We also found that Sp1 can synergistically activate murin RAG2 promoter with Pax-5 or c-Myb. Conclusion Although high homology exist between mouse and human RAG2 promoter, they differs in activity and binding transcriptional factors.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2003年第9期707-711,共5页
Chinese Journal of Microbiology and Immunology
