Part:BBa_K4739003
VgrG3, secretion tip of T6SS
VgrG3 proteins puncture the target cell membrane to deliver the toxic effector molecules. Various VgrG proteins might be associated with different effectors and might target different cells, helping the bacteria to deal with various competitive environments and hosts. In T6SS formation, VgrG3 will form a trimer and puncture target cell when injected from V. cholerae.
The following image shows how T6SS components carry effectors[1]
In our project, we utilize the natural linker by replacing the effector with our designed drugs. Notice that this part contains a linker domain, so subsequent teams can use this part by directly fusing coding part after this part, an example can be viewed at BBa_K4739005. The linker is marked in red in the following structure graph of VgrG3:
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Engineering and Results
We constructed several plasmids based on pBAD33, we display them as follows:
We introduce these plasmids into V. cholerae V52 rhh. We conducted Western blot analysis. We found that our plasmids can be expressed stably since we can detect the signals. However, the induction conditions were not good enough to prevent all cell lysis in the supernatant, we will continue to conduct experiments after wiki freeze.
Reference
1. Cherrak, Y.; Flaugnatti, N.; Durand, E.; Journet, L.; Cascales, E. Structure and Activity of the Type vi Secretion System. Microbiology Spectrum 2019, 7, doi:https://doi.org/10.1128/microbiolspec.psib-0031-2019.
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