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
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 350
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 350
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 350
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 350
- 1000COMPATIBLE WITH RFC[1000]
None |