Experimental strategies for cloning or identifying genes encoding DNA-binding proteins.
This article describes experimental strategies for cloning or identifying genes encoding DNA-binding proteins. DNA-binding proteins are most commonly identified by electrophoretic mobility-shift assay (EMSA) or DNase I footprinting. To identify the gene encoding a protein detected by EMSA or DNase footprinting, the protein often needs to be purified and its sequence analyzed, as described here. Other methods are also available which do not resort to protein purification, including the one-hybrid screen, in vitro expression library screen, and mammalian expression cloning. These methods are outlined, and their advantages and disadvantages are discussed.
Michael F
MF
Carey
Craig L
CL
Peterson
Stephen T
ST
Smale
D003001:Cloning, Molecular / Q000379:methods*; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D011485:Protein Binding; D017422:Sequence Analysis, DNA
D004268:DNA-Binding Proteins