Cloning and Expression of Recombinant Human IGF-I Gene in pET-DsbA

Objective To clone the human insulin like growth factorⅠgene, construct its expression vector and expression it in E.coli.Method The human insulin like growth factorⅠwas amplified from human liver cDNA by RT-PCR and then inserted into the KS vector.IGF-Ⅰ was inserted into pET-DsbA.The recombinant plasmid was identified by enzyme digestion, sequencing and expressed in E.coli.Result IGF-Ⅰ gene in plasmid pET-DsbA-IGF-Ⅰ was successfully expressed in BL21(DE3)plysS.Proteins were identified by SDS-PAGE and Western Blot.Conclusion The recombinant IGF-Ⅰ protein expressed with pET-DsbA-IGF-Ⅰ could be used for the development of its biological products.