Work

Cyclic Nucleotide-Dependent Screening of a λ Expression Library for Nucleic Acid Binding Activities

Year: 2002

Type: article

Abstract: Here we describe a modified ligand screening strategy for the expression cloning of mammalian proteins that require the activation of protein kinase cascades to activate ligand binding activity. The manipulation of prokaryotic signaling pathways by the application of appropriate inhibitors or agonists to the nitrocellulose filter during functional screening of standard bacteriophage cDNA expression libraries can permit detection of activities that would not otherwise be found in their active state. We have applied this strategy to a A expression library to clone a novel renal cDNA that exhibits cAMP-dependent RNA binding activity when subsequently tested by ectopic expression in mammalian cells.

Source: BioTechniques

DOI: 10.2144/02325st02

Authors: Paul Loflin, Julia E. Lever

Institution: The University of Texas Health Science Center at Houston

Language: English

Topic: RNA Research and Splicing

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,415

Funders: National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases