Abstract: Abstract: Objective To construct the fusion expression vector of the genes of enhancement green fluorescent protein (EGFP) and human tissue inhibitor of metalloproteinases1 (hTIMP1). Methord After extracting mRNA from normal human myocardial tissue, open reading frame of hTIMP1 was obtained by nested-PCR. Then the PCR products was cloned into the pEASY-Blunt Simple vector. After identified by sequence analysis, the hTIMP1 ORF gene was cutted by two enzymes and inserted into pEGFP-C1. Then the recombinant plasmid was transformed into the competent cell of E.coliTOP10, and the correct transforments was screened to be identified. Results The PCR products was about 624bp. We compared the sequence of the hTIMP1 ORF gene which inserted into the pEASY-Blunt Simple vector with the ones reported in Genbank, the sequence was not changed. The recombinant plasmid pEGFP-hTIMP1 was also confirmed correct by identification of PCR and double enzymes’ digestion. Conclusion: We constructed fusion expression vector of the genes of EGFP and hTIMP1 successfully and laid primary basis on further reserches.

Key words: EGFP, TIMP1