论文部分内容阅读
目的构建小鼠HMGB1突变型启动子荧光素酶报告基因。方法 PCR扩增小鼠HMGB1启动子DNA,构建小鼠HMGB1野生型启动子荧光素酶报告基因pGL3-HMGB1-Y。重叠延伸PCR突变HSE核心序列,构建HMGB1突变型启动子荧光素酶报告基因pGL3-HMGB1-T,序列比对。结果序列对比结果显示pGL3-HMGB1-Y中HSE核心碱基-TTCGAGAA-已突变为-TACGAGCC-。结论成功构建小鼠HMGB1突变型启动子荧光素酶报告基因,为研究热休克转录因子1对HMGB1的转录调控作用奠定了基础。
Objective To construct a mouse HMGB1 mutant promoter luciferase reporter gene. Methods The mouse HMGB1 promoter DNA was amplified by PCR, and the mouse HMGB1 wild-type luciferase reporter gene pGL3-HMGB1-Y was constructed. The core sequence of HSE was overlapped and extended, and the HMGB1 mutant promoter luciferase reporter gene pGL3-HMGB1-T was constructed. Results The sequence comparison showed that the HSE core-base TTCGAGAA- in pGL3-HMGB1-Y had been mutated to-TACGAGCC-. Conclusion The luciferase reporter gene of mouse HMGB1 mutant promoter was successfully constructed, which lays the foundation for studying the transcriptional regulation of HMGB1 by heat shock transcription factor 1.