title: Purification of biologically active human immunodeficiency virus rev protein from Escherichia coli.
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.
authors.affiliation: Department of Molecular Oncology, Roche Institute of Molecular Biology, Nutley, New Jersey.
authors.forename: A W
authors.initials: AW
authors.lastname: Cochrane
authors.forename: C H
authors.initials: CH
authors.lastname: Chen
authors.forename: R
authors.initials: R
authors.lastname: Kramer
authors.forename: L
authors.initials: L
authors.lastname: Tomchak
authors.forename: C A
authors.initials: CA
authors.lastname: Rosen
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015967:Gene Expression Regulation, Viral; D015694:Gene Products, rev / Q000235:genetics / Q000302:isolation & purification*; D005822:Genetic Vectors; D006678:HIV / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D015534:Trans-Activators / Q000302:isolation & purification*; D014779:Virus Replication / Q000235:genetics; D054321:rev Gene Products, Human Immunodeficiency Virus
chemical_list: D004279:DNA, Viral; D015694:Gene Products, rev; D011994:Recombinant Proteins; D015534:Trans-Activators; D054321:rev Gene Products, Human Immunodeficiency Virus