摘要
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 the luciferase expression vector containing mouse HMGB1 mutant type promoter. Methods The luciferase expression vector containing mouse HMGB1 wild type promoter, pGL3 - HMGB1 - Y, was construct- ed after the fragment of HMGB1 promoter was amplified by PCR. The luciferase reporter gene containing mouse HMGB1 mutant type promoter, pGL3- HMGB1 -T, was constructed after the moUse HMGB1 wild type promoter was mutated by overlap ex- tension PCR. Sequence alignment was conducted between pGL3- HMGB1 - Y and pGL3- HMGB1 - T. Results The se- quence comparision showed that HSE core base - TTCGAGAA- of pGL3 - HMGB1 - Y had been mutated into - TACGAC_~C -. Conclusions The luciferase expression vector containing mouse HMGB1 mutant type promoter, pGL3 - HMGB1 - T, is successfully constructed. It will provide essential material for studying the transcriptional regulation of heat shock transcription factor 1 （HSF1） on HMGB1.
出处
《实用预防医学》
CAS
2012年第8期1127-1129,共3页
Practical Preventive Medicine
基金
国家自然科学基金(30672035)
