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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 신문
유전학
This content is Free Access.
JoVE 신문 유전학
In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
DOI:

09:16 min

March 25, 2020

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Chapters

  • 00:05Introduction
  • 00:49Split-and-Mix Synthesis of Mutagenic Primers
  • 02:34Performing Compartmentalized In Vitro Transcription-translation Reaction
  • 05:07Continued Processing of Libraries and Selection
  • 07:20Results: Examples of Screening of NlaIV Variants for Sequence Specificity Alteration
  • 08:16Conclusion

Summary

자동 번역

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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