Expression of human insulin-like growth factor 1 in eukaryotic cell and detection of its activity
Abstract：Objective To construct eukaryotic expression plasmid for humaninsulin-like growth factor 1(IGF-1) . Methods IGF-1 cDNA was cloned by PCR from the human liver cDNA library， and inserted into eukaryotic expression vector pcDNA3 with gene recombinant technique， forming recombinant pcDNA3-IGF-1 plasmid. The latter was transfected into COS7 cells with liposome-mediated method．Concentration and activity of IGF-1 in the supernatant were detected by ELISA and fibro-blast proliferation assays respectively. Results IGF-1cDNA sequence and its insertion direction in recombinant pcDNA3-IGF-1 plasmid were are correct. The COS7 cells transfected with the pcDNA3-IGF-1 plasmid secreted high concentration of IGF-1，which markedly induced the proliferation of human fibroblast and NIH3T3 cells. Conclusion The recombinant pcDNA3-IGF-1 plasmid can express active IGF-1 with high efficiency， which facilitates the study of its physical and pathological roles in vivo.
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Publication Date：2001-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 1-4 )