Abstract
A method is described to purify recombinant HIV-1 protease from soluble extracts of Escherichia coli. The isolation involves QAE-Sepharose anion exchange chromatography, hexyl agarose hydrophobic interaction chromatography, MonoS cation exchange chromatography, and Superose 6 size exclusion chromatography. Approximately 100 micrograms of protease was obtained from 18 g E. coli paste. The protein was judged to be homogeneous due to the presence of a single band on a silver-stained SDS polyacrylamide gel.
MeSH terms
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Chromatography, Agarose
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Chromatography, Gel
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Chromatography, Ion Exchange
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Cloning, Molecular
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Electrophoresis, Polyacrylamide Gel
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Escherichia coli / genetics
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Gene Products, gag / metabolism
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HIV Protease / chemistry
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HIV Protease / isolation & purification*
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Humans
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Protein Precursors / metabolism
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Sepharose / analogs & derivatives
Substances
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Gene Products, gag
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Protein Precursors
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Recombinant Proteins
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p55 gag precursor protein, Human immunodeficiency virus 1
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aminohexyl-sepharose
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Sepharose
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HIV Protease