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PMID: 3889837 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Efficient secretion and purification of human insulin-like growth factor I with a gene fusion vector in Staphylococci.

Nucleic acids research ·Vol. 13 ·No. 4 ·1985-02-25 ·Pages 1151-62

Nilsson B, Holmgren E, Josephson S, Gatenbeck S, Philipson L, Uhlen M

Abstract

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.

MeSH Terms
Escherichia coli/genetics Gene Expression Regulation Genetic Engineering Genetic Vectors Growth Substances/genetics Hydrolysis Insulin/genetics,isolation & purification,metabolism Insulin Secretion Mutation Peptides/genetics,isolation & purification,metabolism Plasmids Somatomedins/genetics,isolation & purification,metabolism Staphylococcal Protein A/genetics Staphylococcus aureus/genetics
Chemicals
Growth Substances Insulin Peptides Somatomedins Staphylococcal Protein A
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Nilsson B
Holmgren E
Josephson S
Gatenbeck S
Philipson L
Uhlen M
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20 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1985-02-25
Pages
1151-62
Language
English
Region
England
NLM ID
0411011
PMCID
PMC341062
Subset
IM
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