期刊
NATURE METHODS
卷 5, 期 11, 页码 939-942出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nmeth.1262
关键词
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资金
- US National Institutes of Health [81XWH-07-2-0020]
- Defense Threat Reduction Agency [HDTRA1-07-C-0024]
Small libraries for directed evolution can be obtained by neutral drifts that maintain the protein's original function, yielding highly polymorphic, stable and evolvable variants. We describe methods for preparing such libraries, using serum paraoxonase (PON1). An optimized GFP variant fused to PON1 reported levels of soluble, functional enzyme, enabling selection by flow cytometry and identification of enzyme variants exhibiting improved specific and total activities toward several substrates, including toxic organophosphates.
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