Expression in Escherichia coli: purification and properties of the recombinant human general transcription factor rTFIIB.
The human class II transcription factor TFIIB (rTFIIB) was overexpressed in Escherichia coli using a T7 RNA polymerase expression system and further purified to apparent homogeneity. The purified rTFIIB is identical to the endogenous factor according to the following criteria: molecular weight, microsequencing and mass spectra studies, ability to recognize the stable preinitiation complex formed between TFIID and the adenovirus 2 major late TATA box as demonstrated by gel shift as well as by DNase I footprinting assays, and transcription activity.
Moncollin
V
V
Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Faculté de Médecine, Strasbourg, France.
Schaeffer
L
L
Chalut
C
C
Egly
J M
JM
D015153:Blotting, Western; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D010455:Peptides / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification*; D035581:Transcription Factor TFIIB; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*
D010455:Peptides; D011994:Recombinant Proteins; D035581:Transcription Factor TFIIB; D014157:Transcription Factors