Method
Development of a method for screening short-lived proteins using green fluorescent protein
1 Panomics Inc, 2003 East Bayshore Road, Redwood City, CA 94063, USA
2 University of California, San Francisco, Department of Microbiology and Immunology, San Francisco, CA 94143, USA
3 Henry Wellcome Laboratories for Integrative Neuroscience and Endocrinology, Bristol University, Whitson Street, Bristol BS1 3NY, UK
Genome Biology 2004, 5:R81 doi:10.1186/gb-2004-5-10-r81
Published: 28 September 2004Abstract
We have developed a screening technology for the identification of short-lived proteins. A green fluorescent protein (GFP)-fusion cDNA library was generated for monitoring degradation kinetics. Cells expressing a subset of the GFP-cDNA expression library were screened to recover those in which the fluorescence signal diminished rapidly when protein synthesis was inhibited. Thirty clones that met the screening criteria were characterized individually. Twenty-three (73%) proved to have a half-life of 4 hours or less.
