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诱导位点特异性复制遇阻<i>电子。大肠杆菌</i>用荧光阻遏操作员系统
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Biologie
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Journal JoVE Biologie
Inducing a Site Specific Replication Blockage in E. coli Using a Fluorescent Repressor Operator System
DOI:

11:19 min

August 21, 2016

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Chapitres

  • 00:05Titre
  • 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

Traduction automatique

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.

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