Expression of bovine inhibin beta subunit in Escherichia coli.
1. A DNA fragment encoding the beta subunit of bovine inhibin was amplified using the polymerase chain reaction and was cloned in plasmids pUC8 and pUR291. 2. Cultures of Escherichia coli TG2 harbouring pKDK37, a pUR291-derived recombinant plasmid, produced a novel protein with a molecular weight of 130,000 corresponding to a beta-galactosidase-inhibin beta fusion protein. 3. The fusion protein was purified from inclusion bodies by solubilization in 8 M urea followed by an ion-exchange and gel permeation chromatography. 4. Analysis by immunoblotting and competitive radioimmuno assay revealed that the fusion protein was recognized by a monoclonal antibody raised against a chemically synthesized peptide for amino acid residues from +82 to +114 of the beta subunit of the bovine inhibin thereby confirming its identity.
Kim
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KD
Department of Biochemistry and Physiology, University of Reading, Whiteknights, England.
Knight
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PG
Savva
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D028341:Activins; D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle / Q000235:genetics; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded; D004926:Escherichia coli / Q000235:genetics; D007265:Inhibins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D001616:beta-Galactosidase / Q000235:genetics
D000911:Antibodies, Monoclonal; D004277:DNA, Single-Stranded; D011993:Recombinant Fusion Proteins; D028341:Activins; D007265:Inhibins; D001616:beta-Galactosidase