Fast gel electrophoresis to analyze DNA-protein interactions.
A rapid method for electrophoresis of DNA-protein complexes is described. Popular "gel-shift" assays are performed using Pharmacia PhastSystem with its convenience of pre-cast gels and buffer strips and microprocessor-controlled high-resolution separation. Using this system, the products of a DNA binding reaction (DNA-protein complexes) can be separated from "free" DNA in less than one hour. DNA fragments as well as oligonucleotides have been used as targets with partially purified extracts containing sequence-specific DNA binding proteins. We present here a comparison of results of gel-shift assays obtained by conventional techniques and by our rapid "PhastShift" method which reduces the time, effort and technical expertise necessary to obtain reproducible results.
Ramanujam
P
P
Pharmacia P-L Biochemicals, Inc., Milwaukee, WI 53202.
Fogerty
S
S
Heiser
W
W
Jolly
J
J
D000956:Antigens, Viral / Q000378:metabolism; D003201:Computers; D004268:DNA-Binding Proteins / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D019309:Epstein-Barr Virus Nuclear Antigens; D017874:Immediate-Early Proteins; D054334:Viral Regulatory and Accessory Proteins / Q000378:metabolism
D000956:Antigens, Viral; D004268:DNA-Binding Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; D017874:Immediate-Early Proteins; D054334:Viral Regulatory and Accessory Proteins; C048925:herpes simplex virus, type 1 protein ICP4