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Title: The expression of soluble, full-length, recombinant human TSH receptor in a prokaryotic system.
Austin Authors: Busuttil, Bridget E;Turney, K L;Frauman, Albert G 
Affiliation: Clinical Pharmacology and Therapeutics Unit, Department of Medicine, Austin and Repatriation Medical Centre, The University of Melbourne, Austin Campus, Heidelberg, Victoria, 3084, Australia
Issue Date: 1-Dec-2001
Publication information: Protein Expression and Purification; 23(3): 369-73
Abstract: For the first time soluble, full-length, recombinant, human thyroid-stimulating hormone (TSH) receptor (TSHR) has been expressed in a prokaryotic system. The full-length TSHR cDNA, obtained from normal human thyroid, was cloned into a pQE-9 vector, sequenced, and confirmed to be identical to the published sequence, to be full length, and to be in frame. Expression of the receptor was as a fusion protein with a hexahistidine tail at the amino terminal, in an Escherichia coli expression system. Approximately 2.5 mg of protein per liter of bacterial culture was recovered from the cell homogenate, after a single passage through a nickel-nitrilotriacetic acid resin column. An estimated 60% increase in purity of a band of expected size, 87 kDa, was observed upon gel electrophoresis and staining with Coomassie blue, after the single purification step. Immunoreactivity of the 87-kDa protein with Graves' sera was confirmed by Western blotting.
Gov't Doc #: 11722172
DOI: 10.1006/prep.2001.1519
Type: Journal Article
Subjects: Amino Acid Sequence
Blotting, Western
Chromatography, Affinity
Escherichia coli.genetics
Gene Expression
Graves Disease.immunology
Receptors, Thyrotropin.genetics.immunology.isolation & purification.metabolism
Recombinant Fusion Proteins.isolation & purification.metabolism
Appears in Collections:Journal articles

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