Construction and identification of expression vector of human ID4 gene promoter based on bioinformatics analysis

Objective:To construct expression vector of human ID4 gene promoter on the basis of bioinformatics analysis.Methods: According to Internet-based bioinformatics analysis of human ID4 gene promoter structure and transcriptional regulatory elements,the upstream-621bp sequence of ID4 gene transcriptional start site which includes glucocorticoid receptor,cAMP binding protein and estrogen receptor response elements was amplified by PCR.The PCR product of the 621bp sequence was purified and linked to pGEM-T Easy vector by T4 DNA ligase.Recombinant of human ID4 gene promoter-pGEMT Easy was transformed into DH5α competent E.coli.Then the positive recombinant was identified by nested PCR and DNA sequencing.Results and Conclusion:The human ID4 promoter-pGEM-T Easy recombinant plasmid was successfully constructed.The construction of expression vector of human ID4 promoter provides a platform for further researches on promoter activity and expression regulation of human ID4 gene.