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Development of a method for screening short-lived proteins using green fluorescent protein.

ABSTRACT: 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.

SUBMITTER: Jiang X

PROVIDER: S-EPMC545601 | biostudies-literature | 2004

REPOSITORIES: biostudies-literature

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Development of a method for screening short-lived proteins using green fluorescent protein.

Jiang Xin X Coffino Philip P Li Xianqiang X

Genome biology 20040928 10

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 ...[more]

PMID: 15461799

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