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Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach
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JoVE 杂志 遗传学
Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

Determination of the Optimal Chromosomal Location(s) for a DNA Element in Escherichia coli Using a Novel Transposon-mediated Approach

DOI:
10.3791/55946-v

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11:12 min

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September 11, 2017

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, , ,
1Department of Biology, Section for Functional Genomics and Center for Bacterial Stress Response and Persistence (BASP),University of Copenhagen, 2Department of Microbiology and Infection Control,Statens Serum Institut

Chapters

  • 00:05Title
  • 00:48Collection of Transposon Library
  • 02:49Competition Experiment in LB
  • 03:50Southern Blot Analysis to Monitor Competition Over Time
  • 04:59Identification of the Fittest Clones
  • 07:06Flow Cytometry
  • 08:53Results: Transposon-mediated Insertion of DNA Element in E. coli
  • 10:41Conclusion

Summary

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Here, the power of a transposon-mediated random insertion of a non-coding DNA element was used to resolve its optimal chromosomal position.

Tags

Escherichia ColiDNA ElementTransposonChromosomal LocationCompetition ExperimentsWhole Genome SequencingDARS2 RegionElectrocompetent CellsElectroporationKanamycinTransposon Library

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