Human transcription factor USF gene in E. coli purification and in the FAK promoter in a combination of research.docVIP

 PAGE \* MERGEFORMAT 21 Human transcription factor USF gene in E. coli purification and in the FAK promoter in a combination of research 【Abstract】 Objective: Expression in Escherichia coli of Human transcription factor USF (upstream stimulatory factor), and were isolated and purified for further analysis in FAK (focal adhesion kinase) promoter in the DNA binding capacity. Methods: Human Source USF cDNA was cloned into the expression vector pET28a vector, and then the recombinant plasmid was transformed into E. coli BL21 (DE3), induced expression by IPTG with His6  Tag fusion protein His  USF, through the layer of nickel affinity column Analysis of the purified, SDS  PAGE analysis of identification. EMSA analysis of purified protein and FAK promoter DNA binding conditions. Results: His  USF was highly expressed in E. coli, nickel affinity chromatography purified from the high purity of the protein, and can bind to the FAK promoter. Conclusion: The access to high-level expression of high-purity His  USF fusion protein, for the combination of USF and its target gene transcription regulation laid the foundation for further research. 【Key Words】 USF gene expression of focal adhesion kinase protein transcription factor Many important cellular activities by the bHLH type (basic helix  loop  helix) transcription factors are regulated, USF (upstream stimulatory factor) is this type of important transcription factor. It contains helix  loop  helix domain, with dimerization in the form of integrated into the control region of target genes to regulate gene transcription [1]. Its binding sites on target genes containing CACGTG E  box structure. USF was initially as a late promoter of adenovirus trans-activating factor has been found [1], subsequent studies have shown that the distribution of USF in the organization is extensive [2  3], many of the cell gene has also been its regulation [4  8]. By their target genes are mostly reg