A novel method for the purification of the Xenopus transcription factor IIIA.
We have developed a quick and simple purification method that yields large quantities of the Xenopus zinc finger protein transcription factor IIIA (TFIIIA). The protein is purified in the form of the 7S storage particle (TFIIIA/5S RNA complex) found in the ovaries of immature Xenopus. Our method yields 0.5 to 1 mg of pure 7S particle per ovary. It involves a high speed centrifugation step, fractionation on a gel filtration column and a precipitation step using calcium chloride. TFIIIA purified using this protocol retains full DNA binding activity and has the expected zinc ion content.
Miller
J
J
MRC Laboratory of Molecular Biology, Cambridge, UK.
Fairall
L
L
Rhodes
D
D
D000818:Animals; D002850:Chromatography, Gel; D004268:DNA-Binding Proteins / Q000302:isolation & purification*; D005260:Female; D007202:Indicators and Reagents; D008667:Metalloproteins / Q000302:isolation & purification*; D010053:Ovary / Q000378:metabolism*; D012341:RNA, Ribosomal, 5S / Q000302:isolation & purification; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors / Q000302:isolation & purification*; D014982:Xenopus laevis; D015032:Zinc / Q000302:isolation & purification*
D004268:DNA-Binding Proteins; D007202:Indicators and Reagents; D008667:Metalloproteins; D012341:RNA, Ribosomal, 5S; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors; D015032:Zinc