Development of a method for screening short-lived proteins using green fluorescent protein.
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.
Jiang
Xin
X
Panomics Inc, 2003 East Bayshore Road, Redwood City, CA 94063, USA. xjiang@panomics.com
Coffino
Philip
P
Li
Xianqiang
X
D002460:Cell Line; D003001:Cloning, Molecular; D003513:Cycloheximide / Q000494:pharmacology; D005189:False Positive Reactions; D005453:Fluorescence; D015723:Gene Library; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D006207:Half-Life; D006801:Humans; D007700:Kinetics; D014176:Protein Biosynthesis / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011506:Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D013997:Time Factors
D011500:Protein Synthesis Inhibitors; D011506:Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D003513:Cycloheximide