AhpCp2D1 is a ROS-induced promoter controlled by OxyR (transcription factor) which is activated by ROS (Reactive Oxygen Species).
AhpCp2D1 is composed of AhpCp2 (Part:BBa_K1104205), reverse promoter DsbGp (Part:BBa_K1104208),and AhpCp1 (Part:BBa_K1104207). There are also two dual-TFBSs (Transcription Factor Binding Site) for OxyR binding between AhpCp2 and DsbGp.
We improved a BioBrick Part: ahpC promoter (Part:K362001) designed by 2010 KIT-Tokyo team. On PartRegistry, the complex part(according to Ecocyc) composition contains hybrid promoters, shared TFBS (Transcription Factor Binding Site), and reverse promoter DsbG. In this part AhpCp2D1, we succesfully mutated the PstI cutting site (ctgcag->ctacag) of ahpC promoter (Part:K362001), and removed the dsbG coding sequence.
ahpC promoter, as well as its improvement, can be activated by OxyR (Part:BBa_K1104200).
We annotated it thouroughly based on data from (Ecocyc), and found that it contains dsbG coding sequence, AhpCp2 (Part:BBa_K1104205), reverse promoter DsbGp (Part:BBa_K1104208),and AhpCp1 (Part:BBa_K1104207), and a PstI cutting site. Thus we improved the promoter by first mutating the PstI cutting site in ahpCp (Part:BBa_K362001) and make dsbG coding removed.
How ahpC (Part:BBa_K362001) is improved?
In this part,the PstI cutting site in ahpC (BBa_K362001) is mutated at one point, and the Dsbg coding sequence is removed.
Here is the overview about the other ahpC promoter (Part:BBa_K362001) related improvements:
|AhpCp1000 (Part:BBa_K1104203)||The PstI cutting site is mutated.|
|AhpCp2D1 (Part:BBa_K1104204)||After mutating the PstI cutting site, the truncated coding sequence from the DsbG promoter sequence is removed.|
|AhpCp2 (Part:BBa_K1104205)||Only one promoter(AhpCp2) and its TFBS.|
|AhpCpD1 (Part:BBa_K1104206)||Bidirectional promoter: AhpCp1 and DsbGp(reverse promoter), and their shared TFBS.|
|AhpCp1 (Part:BBa_K1104207)||Only one promoter(AhpCp1) and its TFBS.|
Usage and Biology
We designed circuit fighting against Nosema ceranae. After Nosema ceranae infected midgut cells of bees, and Bee. coli should sense the pathogen first before the following circuit(fighting against Nosema ceranae)is triggered, and substance such as Defensin(Part:BBa_K1104300), Abaesin(Part:BBa_K1104301) (more details on Killing Nosema page) in the following circuit will express.
To enhance the strength , we added a device (more details on Sensing Nosema page).
Not only for sensing the midgut oxygen stress caused by Nosemasis, in our project, there are some previous iGEM team who used OxyR-induced promoter, such as 2010 KIT-Tokyo team using it for the ink of E.coli Pen colorful. And those OxyR-induced promoters relative to the projects.
Since ROS is a natural byproduct of the metabolism and have important roles in cell signaling and homeostasis, the improvement of ROS-induced promoter is useful for sensing some specific conditions. ROS is found to involve in cancer, aging and apoptosis. Thus ROS-induced promoter in the future application can be apply to diagnosis and therapy of certain kinds of physical phenomena or disease.
Devasagaya, TPA; Tilak JC, Boloor KK, Sane Ketaki S, Ghaskadbi Saroj S, Lele RD (October 2004). "Free Radicals and Antioxidants in Human Health: Current Status and Future Prospects". Journal of Association of Physicians of India (JAPI) 52: 796.
|AhpCp1000 (Part:BBa_K1104203)||AhpCp2D1 (Part:BBa_K1104204)||AhpCp2 (Part:BBa_K1104205)||AhpCpD1 (Part:BBa_K1104206)||AhpCp1 (Part:BBa_K1104207)||DsbGp (Part:BBa_K1104208)|
|AhpCp1000+E0840 (Part:BBa_K1104243)||AhpCp2D1+E0840 (Part:BBa_K1104244)||AhpCp2+E0840 (Part:BBa_K1104245)||AhpCpD1+E0840 (Part:BBa_K1104246)||AhpCp1+E0840 (Part:BBa_K1104247)||--|
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC