title: Purification and characterization of recombinant equine infectious anemia virus reverse transcriptase.
abstract: A 1.67-kb segment of the equine infectious anemia virus pol gene, encoding a 66-kDa reverse transcriptase (RT), was cloned and expressed in Escherichia coli. Recombinant RT, purified by a combination of metal chelate affinity chromatography and ion-exchange chromatography, displays both RNA-dependent DNA polymerase and RNase H activity. The affinity of purified RT for its replication primer, tRNA(3Lys) was equivalent to that observed for human immunodeficiency virus RT. Our data suggest that an additional domain between RT-RNase H and integrase on the equine infectious anemia virus pol open reading frame is not an integral component of the RT polypeptide.
authors.affiliation: Division of Infectious Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio 44016.
authors.forename: S F
authors.initials: SF
authors.lastname: Le Grice
authors.forename: M
authors.initials: M
authors.lastname: Panin
authors.forename: R C
authors.initials: RC
authors.lastname: Kalayjian
authors.forename: N J
authors.initials: NJ
authors.lastname: Richter
authors.forename: G
authors.initials: G
authors.lastname: Keith
authors.forename: J L
authors.initials: JL
authors.lastname: Darlix
authors.forename: S L
authors.initials: SL
authors.lastname: Payne
mesh_terms: D000818:Animals; D001483:Base Sequence; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D015751:Genes, pol; D006736:Horses; D004860:Infectious Anemia Virus, Equine / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009838:Oligodeoxyribonucleotides; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D016914:Ribonuclease H / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D012194:RNA-Directed DNA Polymerase; D016914:Ribonuclease H