||Improved Method for Screening cDNA Expression Libraries for DNA-Binding Proteins.
||Health Effects Research Lab., Cincinnati, OH. Genetic Toxicology Div.
||c8 Jan 92
DNA-binding proteins ;
Gene library ;
Gene expression ;
Polymerase chain reaction ;
DNA probes ;
DNA polymerase I ;
||Most EPA libraries have a fiche copy filed under the call number shown. Check with individual libraries about paper copy.
The ability to successfully screen a lambda-gt11 complementary DNA expression library for specific gene products that can bind to selected sequences of DNA depends on radioactive double stranded DNA probes with high specific activity. The authors demonstrate here that probes labeled by the polymerase chain reaction are superior to probes made by the Klenow reaction. The use of these PCR-generated probes have facilitated efforts to isolate recombinant phage-containing, putative DNA binding gene products that recognized a 246 base pair transcriptional enhancer region of Rous Sarcoma Virus long terminal repeat.