title: Soluble forms of the human T cell receptor CD4 are efficiently expressed by Streptomyces lividans.
abstract: We have developed a new gene expression and secretion system for Streptomyces lividans and used it to produce soluble forms of a human T-cell receptor CD4 at levels greater than 300 mg/l. The system uses the transcription, translation and secretion signals of the serine protease inhibitor gene STI-II which is naturally produced by S. longisporus. Using these signals, soluble derivatives of CD4 were secreted directly into the culture supernatant as correctly processed soluble, biologically active proteins. High level expression of the CD4 proteins depended on the transcription initiation signal, the amino acid sequence surrounding the signal peptide cleavage site and temporally controlled protease activities. We discuss these results in the context of the potential of this system for producing other eukaryotic proteins in Streptomyces.
authors.affiliation: SmithKline Beecham Pharmaceuticals, Gene Expression Sciences, King of Prussia, PA 19406.
authors.forename: J A
authors.initials: JA
authors.lastname: Fornwald
authors.forename: M J
authors.initials: MJ
authors.lastname: Donovan
authors.forename: R
authors.initials: R
authors.lastname: Gerber
authors.forename: J
authors.initials: J
authors.lastname: Keller
authors.forename: D P
authors.initials: DP
authors.lastname: Taylor
authors.forename: E J
authors.initials: EJ
authors.lastname: Arcuri
authors.forename: M E
authors.initials: ME
authors.lastname: Brawner
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D015704:CD4 Antigens / Q000235:genetics* / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007163:Immunosorbent Techniques; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D011993:Recombinant Fusion Proteins; D012995:Solubility; D013302:Streptomyces / Q000235:genetics*; D014361:Trypsin Inhibitors / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D015704:CD4 Antigens; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D014361:Trypsin Inhibitors; C072839:STI2 protein, Streptomyces; D001616:beta-Galactosidase; D010450:Endopeptidases