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Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis
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Journal JoVE Biologie
Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis

Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis

DOI:
10.3791/51934-v

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13:31 min

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October 31, 2014

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1Department of Biological Sciences, Center for Applied Chemical Biology,Youngstown State University

Chapitres

  • 00:05Titre
  • 02:27Electroporation of Competent Cells
  • 04:35Gridding Transformants and Replica Plating to Determine the Auxotroph Phenotype
  • 08:02Gene Rescue
  • 11:13Results: Generation of Auxotrophic Transformants in Enterobacter sp. YSU
  • 12:39Conclusion

Summary

Traduction automatique

Traduction automatique

Enterobacter sp. YSU grows in glucose minimal salts medium. Auxotrophs are generated by transforming it with a transposome which randomly inserts itself into the host genome. Mutants are found by replica plating from complex medium to minimal medium. Interrupted genes are identified by gene rescue and sequencing.

Tags

Enterobacter Sp. YSUAuxotrophsTransposon MutagenesisTn5 TransposomeM-9 Minimal MediumR6K Gamma Replication OriginKanamycin ResistanceSanger SequencingBLASTGene Identification

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