Differential precipitation and zinc chelate chromatography purification of biologically active HTLV-I Tax1 expressed in E. coli.
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.
Lindholm
P F
PF
Laboratory of Molecular Virology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Marriott
S J
SJ
Gitlin
S D
SD
Brady
J N
JN
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
D002712:Chlorides; D016356:Gene Products, tax; D016328:NF-kappa B; D017967:Zinc Compounds; C016837:zinc chloride; D004247:DNA; D015032:Zinc; D000645:Ammonium Sulfate