Improved method for screening cDNA expression libraries for DNA-binding proteins.

M C Harline; J C Kandala; R D Sage; R V Guntaka; A DeAngelo

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PubMed · 1389172

Improved method for screening cDNA expression libraries for DNA-binding proteins.

M C Harline·J C Kandala·R D Sage·R V Guntaka·A DeAngelo

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Abstract

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.

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Animals Bacteriophage lambda Chickens Cloning, Molecular DNA DNA-Binding Proteins Gene Library Genetic Vectors Plasmids Polymerase Chain Reaction Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.Research Support, U.S. Gov't, P.H.S.

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M C Harline, J C Kandala, R D Sage, R V Guntaka, A DeAngelo. 1992. Improved method for screening cDNA expression libraries for DNA-binding proteins.. https://pubmed.ncbi.nlm.nih.gov/1389172/

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