Recombinant expression, purification and characterisation of the HMG domain of human SRY.
The HMG domain is a DNA binding and bending 'architectural' motif involved in chromatin re-modelling during transcription. Recombinant SRY HMG domain protein, 88 amino acids in length, has been produced in E. coli. Using FPLC and a stirred ultra-filtration cell, this domain has been purified to homogeneity and concentrated to yield milligram quantities. Functional characterisation studies of the pure, concentrated SRY HMG domain show the recombinantly expressed protein to be active in terms of DNA binding and calmodulin binding activities.
Kelly
Sabine
S
Prince Henry's Institute of Medical Research, PO Box 5152, Clayton, Victoria, 3168, Australia.
Yotis
Julia
J
Macris
Mary
M
Harley
Vincent
V
D001665:Binding Sites; D002147:Calmodulin / Q000378:metabolism*; D041322:Chromosomes, Human, Y; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006609:High Mobility Group Proteins / Q000378:metabolism*; D006801:Humans; D008297:Male; D009687:Nuclear Proteins; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D019849:Sex Determination Processes; D051876:Sex-Determining Region Y Protein; D013329:Structure-Activity Relationship; D014157:Transcription Factors
D002147:Calmodulin; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D009687:Nuclear Proteins; D011994:Recombinant Proteins; C497745:SRY protein, human; D051876:Sex-Determining Region Y Protein; D014157:Transcription Factors; D004247:DNA