小鼠热休克转录因子1真核表达载体的构建

Construction of a eukaryotic expression vector containing mouse heat shock transcription factor 1

背景:研究表明,热休克转录因子1具有热休克应答效应。目的:构建小鼠热休克转录因子1(heat shock transcription factor1,HSF1)真核表达载体。方法:克隆小鼠HSF1cDNA,将其插入载体pcDNA3.1,构建热休克转录因子1真核表达载体pcDNA3.1-HSF1。结果与结论:构建的pcDNA3.1-HSF1重组体双酶切电泳后出现大小约5.5kb与1.5kb的2个片段,测序结果显示克隆的小鼠HSF1序列与Genbank中的一致,采用Westernblot法可检测到一条相对分子质量约72000的重组HSF1蛋白条带,说明成功构建小鼠HSF1真核表达载体pcDNA3.1-HSF1。

BACKGROUND： Heat shock transcription factor 1 has heat shock response effect. OBIECTIME： To construct a eukaryotic expression vector containing mouse heat shock transcription factor 1 （HSF1）. METHODS： Amplified mouse HSF1 cDNA fragment was cloned into pcDNA3,1 to construct a eukaryotic expression vector pcDNA3.1-HSFI. RESULTS AND CONCLUSION： There were two fragments in electrophoresis after restriction enzyme digestion, 5.5 kb fragment and 1.5 kb fragment. The result of DNA sequencing showed that the sequence of the cloned HSF1 was identical to that GeneBank had reported. HSF1 protein band with the relative molecular mass of 72 000 was detected by western blot. These findings indicate that the eukaryotic expression vector pcDNA3.1-HSF1 containing mouse HSF1 is successfully constructed.