Capillary electrophoretic mobility shift assay (CEMSA) of a protein-DNA complex.
The utility of capillary electrophoresis in the study of DNA-protein binding is demonstrated, using the minimal DNA binding domain of the onco-protein c-Myb (R2R3) and a specific target DNA sequence as a model system. The capillary electrophoresis method is based on simple UV detection at 260 nm with a linear polymer buffer and a coated capillary, and requires no labeling or derivatization of the DNA. A specific protein-DNA complex is observable as a retarded peak, which increases with increasing protein concentration with a corresponding reduction in the free DNA peak. With DNA and protein preparations of known concentrations, a test for sequence-specific binding can be completed within 10 min.
Xue
B
B
Department of Bioanalysis and Biochemistry, Nycomed Imaging, Oslo, Norway.
Gabrielsen
O S
OS
Myrset
A H
AH
D000818:Animals; D002645:Chickens; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D019075:Electrophoresis, Capillary / Q000379:methods*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D020598:Proto-Oncogene Proteins c-myb; D015534:Trans-Activators / Q000378:metabolism*
D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D020598:Proto-Oncogene Proteins c-myb; D015534:Trans-Activators; D004247:DNA