Efficient secretion and purification of human insulin-like growth factor I with a gene fusion vector in Staphylococci.
A novel approach for production of small polypeptides, using a staphylococcal protein A vector, is described. This system is used to express, secrete and purify human insulin-like growth factor I (IGF-I). A fusion protein consisting of protein A and IGF-I is recovered in high yield by passing the culture medium through an IgG affinity column. Using site-specific mutagenesis an acid labile asp-pro cleavage site was introduced at the fusion point between the two proteins. The protein A "tail" can thereby be removed from the affinity purified fusion protein by chemical cleavage releasing biologically active IGF-I molecules.
D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D005818:Genetic Engineering; D005822:Genetic Vectors; D006133:Growth Substances / Q000235:genetics; D006868:Hydrolysis; D007328:Insulin / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D000078790:Insulin Secretion; D009154:Mutation; D010455:Peptides / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D010957:Plasmids; D013002:Somatomedins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D013205:Staphylococcal Protein A / Q000235:genetics; D013211:Staphylococcus aureus / Q000235:genetics*
D006133:Growth Substances; D007328:Insulin; D010455:Peptides; D013002:Somatomedins; D013205:Staphylococcal Protein A