One plasmid selection system for the rapid evolution of aminoacyl-tRNA synthetases
One plasmid selection system for the rapid evolution of aminoacyl-tRNA synthetases
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DOI：
10.1016/j.bmcl.2009.04.007
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发表时间：
2009-07-15
影响因子：
2.7
通讯作者：
Schultz, Peter G.
中科院分区：
文献类型：
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作者：
Melancon, Charles E., III;Schultz, Peter G.
我们开发了一种快速、直接、单质粒双阳性/阴性选择系统，用于进化大肠杆菌中特异性改变的氨基酰基trna合成酶。该系统利用含有氯霉素乙酰转移酶/尿嘧啶磷酸核糖基转移酶融合基因的琥珀色终止密码子。我们通过从109个变体库中鉴定出一种jannaschmethanococcus tyroyl合成酶变体，证明了该系统的实用性，该变体可以选择性地结合对碘苯丙氨酸来响应琥珀色停止密码子。2009爱思唯尔有限公司版权所有。
We have developed a rapid, straightforward, one plasmid dual positive/negative selection system for the evolution of aminoacyl-tRNA synthetases with altered specificities in Escherichia coli. This system utilizes an amber stop codon containing chloramphenicol acetyltransferase/uracil phosphoribosyltransferase fusion gene. We demonstrate the utility of the system by identifying a variant of the Methanococcus jannaschii tyrosyl synthetase from a library of 109 variants that selectively incorporates para-iodophenylalanine in response to an amber stop codon. (C) 2009 Elsevier Ltd. All rights reserved.