Part:BBa_K4119003

Pvgb-Bs2-Cpa fdx terminator

In this part of our project, the key promoter vgb was a microaerobic induced promoter of Vitreoscilla hemoglobin gene. Considering the gene compatibility difference between different host bacteria, we designed the pMTL-Pvgb-bs2 plasmid to determine whether the promoter vgb could work normally in Clostridium tyrobutyricum by detecting the fluorescent expression intensity of fluorescent protein Bs2.

Results:

The test group: the fluorescence intensity is relatively high

The control group: the fluorescence intensity is lower compared to the test group

The engineered bacteria bearing pMTL-Pvgb-bs2, pMTL-Pthl-bs2 and pMTL82151 plasmids were precultured, inoculated into LB culture medium with different dissolved oxygen, and cultured to OD600 ~0.7 and ~1.2, respectively. The subsequent operation was as recorded by protocol, measuring the fluorescence intensity, collating and the data was analyzed.

The vgb promoter performed a low expression under aerobic conditions, while under microaerobic conditions, it expressed 10.2 folds increased on fluorescence intensity comparing with the constituent promoter, which is exactly as expected.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 350
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 350
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 350
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 350
  
• 1000
  COMPATIBLE WITH RFC[1000]