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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
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JoVE Journal Biology
Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
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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

DOI:
10.3791/2505-v

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09:01 min

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March 16, 2011

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, , , ,
1Department of Microbiology & Environmental Toxicology,University of California Santa Cruz – UCSC

Chapters

  • 00:05Title
  • 01:38Creation of a Mutant Library: Mutagenesis
  • 03:23Creation of a Mutant Library: Readout
  • 04:03Mutant Identification and Characterization: Construction of Gradient
  • 05:06Mutant Identification and Characterization: Stamp Transfer of Bacteria
  • 06:27Mutagenesis and Functional Selection Protocols: Representative Results
  • 08:14Conclusion

Summary

Automatic Translation

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

MutagenesisFunctional SelectionDirected EvolutionProteinsE. ColiMutant LibrariesColE1 PlasmidDNA Polymerase IMutator StrainSolid MediaMutation FrequencyLabelingPCRCloningSequence Space ExplorationBiochemical Activities

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