E. coli, recombinant protein, rhIGF-1, SDS-PAGE, western blot
The hIGF-1 (human Insulin-like Growth Factor 1) is a growth factor produced primarily by the liver under the stimulation of growth hormone (GH). rhIGF-1 is indicated for treatment of primary insulin-like growth factor-1 deficiency (IGFD) and is used for increasing muscle mass by athletes. Many researches have proved that hIGF-1 enhanced glycemic control in Type 1 Diabetes and Type 2 Diabetes. In addition, hIGF-1 has also shown the positive effects in patients with neural disease and cardiovascular disease. Many potential clinical applications of hIGF-1 are still being studied. In order to obtain a large amount of rhIGF-1 for research and treatment, we conduct the research on production of rhIGF-1. In this study, we report results of cloning and expressing rhIGF-1 in E. coli. The higf-1 gene encodes for hIGF-1 was inserted into plasmid pET-22b under the control of T7 promoter to form recombinant plasmid pET22b-IGF1. This expression vector was checked by PCR, restriction enzyme and DNA sequencing. pET22b-IGF1 was transformed into E. coli Origami. The expression of rhIGF-1 was induced by IPTG and confirmed by SDS-PAGE and Western blot using monoclonal anti-hIGF-1 antibody. Overall, we succeeded in expressing rhIGF-1 in E. coli which will be used in the next researches.
Long, D. D., Phuong Thao, D. T., & Phuoc, T. L. (2012). Cloning and expression of hIGF-1 (human Insulin-like Growth Factor 1) in E. coli. Academia Journal of Biology, 34(3se), 78–83. https://doi.org/10.15625/0866-7160/v34n3se.1776