Part:BBa_K4119010

Plac->ΔperR->self-target->Cpa fdx terminator

it's Plac->ΔperR->self-target->Cpa fdx terminator

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 368
  Illegal PstI site found at 1597
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 368
  Illegal PstI site found at 1597
  Illegal NotI site found at 3487
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 368
  Illegal BglII site found at 428
  Illegal XhoI site found at 3359
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 368
  Illegal PstI site found at 1597
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 368
  Illegal PstI site found at 1597
  Illegal NgoMIV site found at 3791
  
• 1000
  COMPATIBLE WITH RFC[1000]

Results:

Fig-2 Plasmid-self-target bacteria colonies PCR results

The results show that the target plasmid colonies PCR results appear band to eliminate the plasmid bacteria no band, at the same time, eliminate the target plasmid bacteria cannot survive in the medium containing resistance, prove that the target bacteria has completed plasmid elimination.

We hope to induce the elimination of plasmids by tetracycline after lactose-induced knockout of the gene of interest, and at present, we have only achieved lactose-induced knockout of the gene of interest with success. In addition, we designed plasmids solely to eliminate plasmids to verify whether the method of eliminating plasmids using tetr is feasible in Clostridium butyrate. We introduced a plasmid carrying the tetR gene in Clostridium butyrate and successfully eliminated it by tetracycline induction, demonstrating that this method is feasible. The work of knocking out the gene of interest and then eliminating the plasmid for time reasons is not completed.