STABLE: protein‐DNA fusion system for screening of combinatorial protein libraries in vitro

FEBS Letters

Doi, Nobuhide¹  Yanagawa, Hiroshi¹ 

[1]Mitsubishi Kasei Institute of Life Sciences, 11 Minamiooya, Machida, Tokyo 194-8511, Japan

关键词: In vitro transcription/translation;    Water in oil emulsion;    Streptavidin-biotin complex;    Directed protein evolution;    In vitro selection;    cDNA library;    FITC;    fluorescein isothiocyanate;    PAGE;    polyacrylamide gel electrophoresis;    PCR;    polymerase chain reaction;    STA;    streptavidin;   

DOI  :  10.1016/S0014-5793(99)01041-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have developed a new method that permits the complete in vitro construction and selection of peptide or protein libraries. This method relies on an in vitro transcription/translation reaction compartmentalized in water in oil emulsions. In each emulsion compartment, streptavidin (STA)-fused polypeptides are synthesized and attached to the encoding DNA via its biotin label. The resulting protein-DNA fusion molecules recovered from the emulsion can be subjected to affinity selection based on the properties of the peptide portion, whose sequence can be determined from that of its DNA-tag. This method, named ‘STABLE’ (STA-biotin linkage in emulsions), should be useful for rapid in vitro evolution of proteins and for ligand-based selection of cDNA libraries.

【 授权许可】

Unknown