Abstract: Purpose: To further study the effect of E 2 F 1 transcription factor on the transcriptional activation of β promoter, and explain the transcriptional regulation mechanism on MTS 1 gene. Methods:The recombinant plasmid containing 2 or 3 mutant or deleted on E 2 F 1 A, B, C-bound locus sequence on the 0.38 kb fragment cut by SacⅡand SacⅠ of the β promoter was constructed by PCR site-directed mutagenesis and enzymatic cutting and ligating. The recombinant plasmids containing 2 or 3 mutant or deleted on E 2 F 1 A, B, C-bound locus sequence were transfected into Jurkat cells, which were biallelic deletion of MTS 1 gene by transient transfection. Luciferase report gene was used to observe β promoter transcriptional activation. Results: Four new recombinant plasmids containing the mutant with two bound loci of E 2 F 1 A,B,C were obtained separately by PCR, and a recombinant plasmid containing all the three mutant on locus was constructed by enzymatic cutting and ligating, and identified by SacⅠ or NaeⅠenzymatic cutting, and sequencing. The luciferase expression of recombinant plasmid in Jurkat cells decreased, especially the mutant 3 bound locus sequence of E 2 F 1 A, B, C, as compared to the wild-type recombinant plasmid on E 2 F 1-bound locus sequence. Conclusion: These newly-constructed recombinant plasmids can be used to study the function of the transcriptional activation of MTS 1 gene by gene transfection. There is a potential relation between the transcriptional activation of MTS 1 gene β promoter and the transactivation of E 2 F 1 transcription factor.

Key words: MTS 1 gene, E 2 F 1 transcriptional factor, Gene mutation