Improved method for screening cDNA expression libraries for DNA-binding proteins.
The ability to successfully screen a lambda gt11 cDNA 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. We demonstrate here that probes labeled by the PCR are superior to probes made by the Klenow reaction. The use of these PCR-generated probes have facilitated our 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.
Harline
M C
MC
Department of Molecular Microbiology and Immunology, University of Missouri-Columbia 65212.
Kandala
J C
JC
Sage
R D
RD
Guntaka
R V
RV
DeAngelo
A
A
D000818:Animals; D010582:Bacteriophage lambda / Q000235:genetics; D002645:Chickens; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D015723:Gene Library; D005822:Genetic Vectors; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*
D004268:DNA-Binding Proteins; D004247:DNA