Examination of alternate codon bias solutions for expression and purification of recombinant mechano‐growth factor in Escherichia coli

• DOI: 10.1002/bab.1312
• Publication Date: 2014-10-27T09:51:31Z
• Authors: Jianguo Feng; Rongxue Wan; Qian Yi; Ling He; Li Yang; Liling Tang
• Keywords: 0301 basic medicine; Base Sequence; Molecular Sequence Data; Gene Expression; Protein Engineering; Recombinant Proteins; Mice; 03 medical and health sciences; Escherichia coli; Mutagenesis, Site-Directed; Animals; Humans; Insulin-Like Growth Factor I; Codon; Cell Proliferation

Abstract Mechano‐growth factor (MGF), an alternative splicing variant of insulin‐like growth factor‐1 ( IGF‐1 ) gene, promotes cell proliferation and inhibits differentiation. It also plays important role in tumor development. is to optimize the production process achieve MGF protein because there no commercial available. In this study, human gene cloned into pGEX‐4T‐1 recombinant (rhMGF) could be expressed Rosetta (DE3) by isopropyl β‐ d ‐1‐thiogalactopyranoside induction but not BL21 (DE3). Mutation from rare codons Escherichia coli preferred ones performed. We obtain MGF(Mut54–56) MGF(Mut‐total) fragments through site‐directed mutagenesis overlapping PCR. Both pGEX‐4T‐1/MGF(Mut54–56)‐ pGEX‐4T‐1/MGF(Mut‐total)‐transformed can induced express rhMGF protein. To technology, expression purification are analyzed compared Results indicate that significantly higher than The yield pGEX‐4T‐1/MGF(Mut‐total) pGEX‐4T‐1/MGF(Mut54–56). test biological activity purified affinity chromatography C2C12 line find significantly. conclusion, we establish a method produce economically with high