Purification of biologically active human immunodeficiency virus rev protein from Escherichia coli.
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.
Cochrane
A W
AW
Department of Molecular Oncology, Roche Institute of Molecular Biology, Nutley, New Jersey.
Chen
C H
CH
Kramer
R
R
Tomchak
L
L
Rosen
C A
CA
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
D004279:DNA, Viral; D015694:Gene Products, rev; D011994:Recombinant Proteins; D015534:Trans-Activators; D054321:rev Gene Products, Human Immunodeficiency Virus