Automated sequential affinity chromatography of sea urchin embryo DNA binding proteins.
An automated method of running a tandem sequence of oligonucleotide affinity columns was used to purify factors that interact specifically with cis-regulatory sites of the CyIIIa cytoskeletal actin gene of the sea urchin embryo (Strongylocentrotus purpuratus). The method allows quantitative enrichment in a single chromatographic run of up to 12 different sequence-specific DNA binding proteins, each of which may then be readily purified to homogeneity by methods such as preparative gel electrophoresis. The affinity chromatography and identification of six different CyIIIa-regulatory factors is described, and the general utility of the method is discussed.
Coffman
J A
JA
Division of Biology, California Institute of Technology, Pasadena 91125.
Moore
J G
JG
Calzone
F J
FJ
Britten
R J
RJ
Hood
L E
LE
Davidson
E H
EH
D000199:Actins / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002846:Chromatography, Affinity; D004268:DNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009838:Oligodeoxyribonucleotides; D012617:Sea Urchins / Q000196:embryology
D000199:Actins; D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides