Abstract
A genetic approach was used to facilitate purification of human immunodeficiency virus (HIV) rev protein. A recombinant protein containing a stretch of six histidine residues at the amino terminus was engineered and overexpressed in Escherichia coli. Purification of greater than 95% was achieved in a single step using an immobilized metal ion chromatography with a resin that has selectivity for proteins with neighboring histidine residues. We show that the modified protein is both properly modified and biologically active.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Cloning, Molecular
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DNA, Viral / genetics
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Escherichia coli / genetics
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Gene Expression Regulation, Viral
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Gene Products, rev / genetics
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Gene Products, rev / isolation & purification*
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Genetic Vectors
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HIV / genetics*
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HIV / physiology
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Humans
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Molecular Sequence Data
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Trans-Activators / isolation & purification*
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Virus Replication / genetics
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rev Gene Products, Human Immunodeficiency Virus
Substances
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DNA, Viral
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Gene Products, rev
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Recombinant Proteins
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Trans-Activators
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rev Gene Products, Human Immunodeficiency Virus