title: Differential precipitation and zinc chelate chromatography purification of biologically active HTLV-I Tax1 expressed in E. coli.
abstract: A protocol which involves sequential ammonium sulfate precipitation and zinc chelate chromatography to purify the HTLV-I Tax1 protein expressed in E. coli is described. The final Tax1 product is greater than 90% pure and the yield is approximately 1 mg per liter of liquid culture. The purified Tax1 protein is biologically active in indirect in vitro DNA binding assays and cellular NF-kB induction experiments.
authors.affiliation: Laboratory of Molecular Virology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
authors.forename: P F
authors.initials: PF
authors.lastname: Lindholm
authors.forename: S J
authors.initials: SJ
authors.lastname: Marriott
authors.forename: S D
authors.initials: SD
authors.lastname: Gitlin
authors.forename: J N
authors.initials: JN
authors.lastname: Brady
mesh_terms: D000645:Ammonium Sulfate; D011232:Chemical Precipitation; D002712:Chlorides; D002846:Chromatography, Affinity / Q000379:methods; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D016356:Gene Products, tax / Q000235:genetics / Q000302:isolation & purification*; D006367:HeLa Cells; D015368:Human T-lymphotropic virus 1 / Q000032:analysis*; D006801:Humans; D016328:NF-kappa B / Q000096:biosynthesis / Q000235:genetics; D015032:Zinc; D017967:Zinc Compounds
chemical_list: D002712:Chlorides; D016356:Gene Products, tax; D016328:NF-kappa B; D017967:Zinc Compounds; C016837:zinc chloride; D004247:DNA; D015032:Zinc; D000645:Ammonium Sulfate