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无核酸酶的表达和纯化原氧基质 3，4-二氧酶
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Biochimica
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JoVE Journal Biochimica
Expression and Purification of Nuclease-Free Oxygen Scavenger Protocatechuate 3,4-Dioxygenase
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无核酸酶的表达和纯化原氧基质 3，4-二氧酶

DOI:
10.3791/59599-v

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10:14 min

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November 08, 2019

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, , ,
1Cancer Biology and Genetics, College of Medicine,The Ohio State University

Capitoli

  • 00:04Titolo
  • 00:25Inducing PCD (protocatechuate 3,4 dioxygenase) Expression in E. coli
  • 03:39Nickel Affinity Chromatography Purification of PCD
  • 04:46Nuclease Activity Assay
  • 06:19PCA (Protocatechuic Acid) Oxidation and Nuclease Activity Assays
  • 08:01Results: Purification of PCD
  • 09:23Conclusion

Summary

Traduzione automatica

Traduzione automatica

原酸3，4-二氧酶（PCD）可以酶分从水系统中分离游离的游离双原子氧，使用其基质原酸（PCA）。该协议描述了这种氧气清除酶的表达、纯化和活性分析。

Tags

Nuclease-freeOxygen ScavengerProtocatechuate 34-dioxygenasePCD ExpressionE. Coli BL21TransformationLB-ampicillin AgarProtein ExpressionIPTGAmmonium Iron SulfateCentrifugationSDS-PAGE

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