STABLE: protein-DNA fusion system for screening of combinatorial protein libraries in vitro
STABLE: protein-DNA fusion system for screening of combinatorial protein libraries in vitro
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DOI：
10.1016/s0014-5793(99)01041-8
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发表时间：
1999-08-27
期刊：
影响因子：
3.5
通讯作者：
Yanagawa, H
中科院分区：
文献类型：
--
作者：
Doi, N;Yanagawa, H
我们已经开发了一种新的方法，允许完整的体外构建和选择的肽或蛋白质文库。该方法依赖于在油包水乳液中区室化的体外转录/翻译反应。在每个乳液隔室中，合成链霉亲和素（STA）融合多肽，并通过其生物素标记物连接到编码DNA上。从乳液中回收的蛋白质-DNA融合分子可以进行基于肽部分的性质的亲和选择，其序列可以由其DNA-标签的序列确定。这种称为“STABLE”（乳液中的STA-生物素连接）的方法应该可用于蛋白质的快速体外进化和cDNA文库的基于配体的选择。(C)1999年欧洲生物化学学会联合会。
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. (C) 1999 Federation of European Biochemical Societies.