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具有更严格特异性的限制内核内核酶的体外定向进化
JoVE Journal
Genética
This content is Free Access.
JoVE Journal Genética
In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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具有更严格特异性的限制内核内核酶的体外定向进化

6,891 Views

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09:16 min

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March 25, 2020

DOI:

10.3791/60807-v

DOI:
10.3791/60807-v

•

09:16 min
March 25, 2020

•

1 Views
,
1International Institute of Molecular and Cell Biology, Warsaw (IIMCB)

Summary

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具有新序列特异性的限制内联酶可以从识别部分退化序列的酶中开发。在这里，我们提供了一个详细的协议，我们成功地用于改变NlaIV酶的序列特异性。该协议的关键成分是转录/翻译反应的体外分块和具有新序列特性的变体的选择。

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Cite this Article

Skowronek, K. J., Bochtler, M. In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity. J. Vis. Exp. (157), e60807, doi:10.3791/60807 (2020).

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