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导演蛋白质的演化突变和功能选择协议
E。大肠杆菌</em
JoVE Journal
Biology
This content is Free Access.
JoVE Journal
Biology
Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in
E. coli
Please note that all translations are automatically generated.
Click here for the English version.
导演蛋白质的演化突变和功能选择协议
E。大肠杆菌</em
DOI:
10.3791/2505-v
•
09:01 min
•
March 16, 2011
•
Chris Troll
,
David Alexander
,
Jennifer Allen
,
Jacob Marquette
,
Manel Camps
1
Department of Microbiology & Environmental Toxicology
,
University of California Santa Cruz – UCSC
Chapters
00:05
Title
01:38
Creation of a Mutant Library: Mutagenesis
03:23
Creation of a Mutant Library: Readout
04:03
Mutant Identification and Characterization: Construction of Gradient
05:06
Mutant Identification and Characterization: Stamp Transfer of Bacteria
06:27
Mutagenesis and Functional Selection Protocols: Representative Results
08:14
Conclusion
Summary
Automatic Translation
English (Original)
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Automatic Translation
在这里,我们展示了一个简单的协议来创建一个给定的目标序列的随机突变体库。我们表明,这种方法,这是在大肠杆菌体内进行,如何可以再加上功能选择进化出新的酶活动。
Tags
Mutagenesis
Functional Selection
Directed Evolution
Proteins
E. Coli
Mutant Libraries
ColE1 Plasmid
DNA Polymerase I
Mutator Strain
Solid Media
Mutation Frequency
Labeling
PCR
Cloning
Sequence Space Exploration
Biochemical Activities
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