title: Development of a vector system for the expression of bioengineered proteins.
abstract: The low natural abundance of many proteins is a major factor in preventing their development as therapeutic or diagnostic tools. To circumvent this barrier, we have used synthetic oligonucleotide technology to construct a gene based on the sequence of a cDNA for human interleukin 6 (IL-6). The synthetic gene encodes a cysteine-free, bioengineered rIL-6 protein, which is expressed at high concentrations in Escherichia coli as a tripartite fusion protein. Cleavage of the fusion protein with collagenase (EC 3.4.24.8) releases a 23-kDa rIL-6 protein that can be easily purified to homogeneity. This rIL-6 protein displays a range of biological activities similar to those of natural human IL-6, as demonstrated by its ability to (a) protect cells from viral infection and (b) stimulate the synthesis of fibrinogen in rat FAZA cells.
authors.affiliation: Department of Pathology, University of North Carolina, Chapel Hill 27599.
authors.forename: J N
authors.initials: JN
authors.lastname: Snouwaert
authors.forename: R C
authors.initials: RC
authors.lastname: Jambou
authors.forename: J E
authors.initials: JE
authors.lastname: Skonier
authors.forename: K
authors.initials: K
authors.lastname: Earnhardt
authors.forename: J R
authors.initials: JR
authors.lastname: Stebbins
authors.forename: D M
authors.initials: DM
authors.lastname: Fowlkes
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D015850:Interleukin-6; D007378:Interleukins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology; D010948:Viral Plaque Assay
chemical_list: D015850:Interleukin-6; D007378:Interleukins; D011993:Recombinant Fusion Proteins