Secretion in Escherichia coli and phage-display of recombinant insulin-like growth factor binding protein-2

Journal of Biotechnology
M R LucicG Forsberg

Abstract

Insulin-like growth factors (IGFs) promote cell growth and differentiation. Their actions are regulated by six different, but related, binding proteins (IGFBPs). To investigate the molecular interactions between IGFs and IGFBPs, an Escherichia coli based production method and a phage display system has been developed. The cDNA for bovine IGFBP-2 was inserted between regions coding for the pelB signal sequence and geneIII product, g3p, of bacteriophage fd in a phagemid vector to generate pGF14. The coding sequences of IGFBP-2 and g3p were separated by an amber stop codon and a flexible linker containing the cleavage recognition site for H64A subtilisin. Using this system in BL21, a non-supE strain lacking ompT, most product, approximately 4 mg 1(-1) of IGFBP-2, was obtained in the growth medium. The bacterially derived IGFBP-2 had a correct N-terminal sequence, molecular mass on SDS-PAGE and the same affinity for IGF-1 and IGF-II as IGFBP-2 from mammalian cells. In a supE strain of E. coli, IGFBP-2 was produced as an IGF-binding fusion to g3p. Procedures for display and approximately 10000 fold enrichment of IGFBP-2 bearing phage using adsorption to IGF-II coated microtitre plates were developed. Thus IGFBP-2 can be secreted in ...Continue Reading

References

May 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·C F BarbasR A Lerner
Aug 1, 1990·Journal of Molecular Endocrinology·F Z UptonF J Ballard
Sep 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·C F BarbasS J Benkovic
Aug 15, 1991·Nature·T ClacksonG Winter
Nov 12, 1991·Biochemistry·H B LowmanJ A Wells
Nov 26, 1986·Biochemical and Biophysical Research Communications·M L HuhtalaM Seppälä
Nov 1, 1989·Endocrinology·S M DonovanR G Rosenfeld
Jan 1, 1989·Proteins·P CarterJ A Wells
Apr 20, 1967·Biochemical and Biophysical Research Communications·I Schechter, A Berger
Jan 1, 1993·Vitamins and Hormones·M M Rechler
May 1, 1994·Trends in Biotechnology·T Clackson, J A Wells
Jan 1, 1995·Protein Engineering·A Knappik, A Plückthun
Dec 5, 1993·Journal of Molecular Biology·H B Lowman, J A Wells
Jul 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·C F BarbasR A Lerner
Oct 1, 1996·Current Opinion in Biotechnology·M R Eckart, C M Bussineau

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Citations

Jan 10, 2008·Applied Microbiology and Biotechnology·Dorra Zouari AyadiSamir Bejar
Oct 24, 2007·Molecular Biotechnology·Xiaobo LiangLiandong Huan
May 26, 2001·Applied and Environmental Microbiology·J ManosroiA Manosroi
Dec 7, 2010·Protein Expression and Purification·Erik VernetMichael Sundström
Sep 17, 2009·Molecular Systems Biology·Daniel M WidmaierChristopher A Voigt
Aug 13, 2011·Biotechnology Journal·Elisabeth LintonCharles D Miller
Apr 23, 2008·Current Protocols in Neuroscience·A Bradbury

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