Abstract:

BACKGROUND: Studies showed that heart transcription factor GATA bind protein 4 (GATA4) are closely related to congenital heart disease. However, there are few reports about the construction of GATA4 expression plasmid and how it effects the development and proliferation of myocardial cell.
OBJECTIVE: To construct the recombinant plasmid pCMV-Myc-GATA4.
METHODS: Recombinant plasmid pUC57-GATA4 was digested by EcoRⅠand Kpn I to obtain the target gene GATA4. Double enzyme digestion was conducted for pCMV-Myc and GATA4. Both fragments were connected by using T4 ligase and transferred to DH5α. Then plasmid was extracted and detected by PCR and double enzyme digestion and sequencing.
RESULTS AND CONCLUSION: The coding area of GATA4 for approximate 1.3 kb was obtained. The results of PCR, enzyme digestion and gene sequencing showed that the recombinant expression plasmid had correct codogenic gene fragment. The recombinant expression plasmid of GATA4 gene is successfully constructed and identified.