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Indusere en Stedsspesifikk Replication Blokkering i<i> E. coli</i> Ved hjelp av en fluoriserende Repressor Operator System
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Biologia
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JoVE Journal Biologia
Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System

Indusere en Stedsspesifikk Replication Blokkering i<i> E. coli</i> Ved hjelp av en fluoriserende Repressor Operator System

DOI:
10.3791/54434-v

•

11:19 min

•

August 21, 2016

•

, , , , ,
1School of Environmental and Life Sciences,University of Newcastle

Capítulos

  • 00:05Título
  • 00:56Blocking Replication with FROS
  • 02:32Fluorescence Microscopy
  • 04:00DNA Extraction/Agarose Plugs
  • 06:17Neutral-neutral 2-Dimensional Gel Electrophoresis
  • 09:22Results: FROS Creates a Site Specific and Reversible Replecation Blockage
  • 10:53Conclusion

Summary

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Tadução automática

We describe here a system utilizing a site-specific, reversible in vivo protein block to stall and collapse replication forks in Escherichia coli. The establishment of the replication block is evaluated by fluorescence microscopy and neutral-neutral 2-dimensional agarose gel electrophoresis is used to visualize replication intermediates.

Tags

E. ColiDNA ReplicationReplication ForkDNA RepairFluorescent RepressorTetR-YFPTetracycline OperatorReplication BlockageTwo-dimensional Gel ElectrophoresisAgarose Pad

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