Construction of recombinant plasmid pEGFP-N1-IGF-1 in rats

Objective To construct the recombinant plasmid pEGFP-N1-IGF-1 for spinal cord injury (SCI) with gene therapy.Methods The cDNA encoding the mature rat insulin-like growth factor 1 was isolated by using RT-PCR method with total RNA extracted from rat liver.IGF-1 gene was cloned into eukaryotic expression vector pEGFP-N1 of EGFP reported gene encoding green fluorescence protein in the form of fusion protein.Results [WTBZ]The cDNA encoding the mature rat insulin-like growth factor-1 (IGF-1) was successfully isolated by using RT-PCR method with total RNA extracted from rat liver.IGF-1 gene was cloned into eukaryotic expression vector pEGFP-N1 of EGFP reported gene encoding green fluorescence protein in the form of fusion protein.IGF-1 gene in the recombinant plasmid pEGFP-N1-IGF-1 was exactly confirmed with DNA sequencing and DNA electrophoresis after enzyme-incise.Conclusion The expression vector of recombinant plasmid pEGFP-N1-IGF-1 is successfully constructed,and could play an important role in gene therrapy of SCI.