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Expression in Escherichia coli of a Chemically Synthesized Gene for Biologically Active Bovine Acidic Fibroblast Growth Factor

Bio/Technology volume 5, pages 960–965 (1987)Cite this article

Abstract

A gene encoding bovine acidic fibroblast growth factor has been chemically synthesized, cloned and expressed as a biologically active protein in Escherichia coli. The 440 base pair gene was assembled by enzymatic ligation of 16 oligonucleotides and cloned into a pBR322–derived expression plasmid downstream of the hybrid tac promoter. Expressed recombinant mitogen comigrated with the native bovine brain–derived protein as detected by Western blot immunological analysis. The expression product of the synthetic gene has been purified to apparent homogeneity and demonstrates a mitogenic activity for Balb/c 3T3 cells in the presence of heparin equivalent to the brain–derived mitogen. These results demonstrate the feasibility of expressing large amounts of functional acidic fibroblast growth factor in bacteria and provide a system for site–specific mutagenesis of the protein.

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  1. David L. Linemeyer: Corrresponding author.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Merck Institute for Therapeutic Research, Merck Sharp and Dohme Research Laboratories, P.O. Box 2000, Rahway, New Jersey, 07065

    David L. Linemeyer, Linda J. Kelly, John G. Menke, Guillermo Gimenez-Gallego, Jerry DiSalvo & Kenneth A. Thomas

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  1. David L. Linemeyer
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Linemeyer, D., Kelly, L., Menke, J. et al. Expression in Escherichia coli of a Chemically Synthesized Gene for Biologically Active Bovine Acidic Fibroblast Growth Factor. Nat Biotechnol 5, 960–965 (1987). https://doi.org/10.1038/nbt0987-960

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