In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
Title:
In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
Author:
Skowronek, Krzysztof J.
Personal Author:
Publication Information:
Cambridge, MA MyJoVE Corp 2016
Physical Description:
online resource (556 seconds)
Series:
Genetics
General Note:
Title from resource description page
Abstract:
Restriction endonucleases with new sequence specificity can be developed from enzymes recognizing a partially degenerate sequence. Here we provide a detailed protocol that we successfully used to alter the sequence specificity of NlaIV enzyme. Key ingredients of the protocol are the in vitro compartmentalization of the transcription/translation reaction and selection of variants with new sequence specificities.
Reading Level:
For undergraduate, graduate, and professional students
Subject Term:
Electronic Access:
https://www.jove.com/t/60807