Abstract
An open reading frame of the alpha-subunit 1-205 residues (alpha205) of human acetylcholine receptor (AchR) was amplified by PCR with pUC-AChR alpha205 as the template and inserted into vector pMAL-c2X. The constructed pMAR alpha205 was transferred into E. coli BL21 which were then grown in LB medium. The amount of soluble MBP-AChR alpha205 protein reached about 25% of total soluble proteins from the cell lysate. Using amylose-affinity chromatography, about 35 mg MBP-AChR alpha205 could be obtained from 1 l culture. Western blot analysis and ELISA showed that immunoreactivities of both MBP-AChR alpha205 and AChR alpha205 were similar to that of AChR alpha-subunit from Torpedo.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Binding Sites
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Carrier Proteins / biosynthesis*
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Carrier Proteins / chemistry*
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Carrier Proteins / genetics
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Carrier Proteins / isolation & purification
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Chromatography, Affinity
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Cysteine Endopeptidases / chemistry*
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Humans
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Maltose-Binding Proteins
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Neoplasm Proteins / chemistry*
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Protein Binding
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Protein Engineering / methods*
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Protein Structure, Tertiary
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Receptors, Nicotinic / biosynthesis*
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Receptors, Nicotinic / chemistry*
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Receptors, Nicotinic / genetics
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Receptors, Nicotinic / isolation & purification
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / isolation & purification
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Solubility
Substances
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Carrier Proteins
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Maltose-Binding Proteins
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Neoplasm Proteins
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Receptors, Nicotinic
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Recombinant Fusion Proteins
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Cysteine Endopeptidases
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cancer procoagulant