Regulatory

Part:BBa_K1104204

Designed by: Ting-Yun Chiang   Group: iGEM13_NYMU-Taipei   (2013-09-16)
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AhpCp2D1

AhpCp2D1 is a ROS-induced promoter controlled by OxyR (transcription factor) which is activated by ROS (Reactive Oxygen Species).

Mutation of ahpC promoter(Part:K362001)

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.

Improvement

We improved a BioBrick Part: ahpC promoter (Part:K362001) designed by [http://2010.igem.org/Team:KIT-Kyoto/Parts 2010 KIT-Tokyo team]. On PartRegistry, the complex part(according to [http://ecocyc.org/ECOLI/new-image?object=EG11384 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 ([http://ecocyc.org/ECOLI/new-image?object=EG11384 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 [http://2013.igem.org/Team:NYMU-Taipei/Project/Inhibition/Killing Killing Nosema] page) in the following circuit will express.

To enhance the strength , we added a device (more details on [http://2013.igem.org/Team:NYMU-Taipei/Project/Inhibition/Sensor Sensing Nosema] page).

Strenthening device

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 [http://2010.igem.org/Team:KIT-Kyoto/Parts 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.

Related Parts

ahpC(Part:BBa_K362001)
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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None