mRNA display for the selection and evolution of enzymes from in vitro-translated protein libraries
mRNA display for the selection and evolution of enzymes from in vitro-translated protein libraries
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DOI：
10.1038/nprot.2011.312
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发表时间：
2011-04-01
期刊：
影响因子：
14.8
通讯作者：
Seelig, Burckhard
中科院分区：
文献类型：
--
作者：
Seelig, Burckhard
MRNA展示技术能够在单个试管中从超过10(12)个不同突变体的文库中进行体外选择和定向进化。这些文库的大小远远超出了筛选技术和大多数体内和体外选择方法的限制。MRNA展示技术已被用于选择与特定配体结合的多肽和蛋白质，以及新的酶。该方案详细说明了产生mRNA展示蛋白(3d)的过程，并使它们经受形成键反应的酶的选择和进化(4-10周)。新的RNA连接酶的产生证明了这种方法的有效性。
The mRNA display technology enables the in vitro selection and directed evolution of functional proteins from libraries of more than 10(12) different mutants in a single test tube. The size of these libraries is well beyond the limit of screening technologies and of most in vivo and in vitro selection methods. The mRNA display technology has been used to select peptides and proteins that bind to a specific ligand, as well as novel enzymes. This protocol details the procedure to produce mRNA-displayed proteins (3 d) and to subject them to a selection and evolution of enzymes for bond-forming reactions (4-10 weeks). This method is demonstrated by the generation of new RNA ligase enzymes.