Difference between revisions of "Part:BBa K1104203"

Revision as of 22:10, 4 October 2013

AhpCp1000

AhpCp1000 is a ROS-induced promoter controlled by OxyR (transcription factor) which is activated by ROS (Reactive Oxygen Species).

AhpCp1000 is composed of part of dsbG coding sequence, 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 (Part:BBa_K1104205), DsbGp (Part:BBa_K1104208)and DsbGp (Part:BBa_K1104208), AhpCp1 (Part:BBa_K1104207).

Mutation of ahpC promoter(Part:K362001)

OxyR is activator of AhpCp1000 promoter. More details about OxyR can be found on the PartRegistry page: Part:BBa_K1104200.

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 AhpCp1000, we succesfully mutated the PstI cutting site (ctgcag->ctacag) of ahpC promoter (Part:K362001).

ahpC promoter, as well as its improvement, can be activated by OxyR (Part:BBa_K1104200).

How is this promoter improved with respect to ahpCp?

In this part,the PstI cutting site in ahpC (K362001) is mutated at one point.

How ahpCp (Part:BBa_K362001) is improved?

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 AhpCp1000.

Here is the overview about the other ahpC promoter (Part:BBa_K362001) improvements:

• 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.
• DsbGp (Part:BBa_K1104208): Only the reverse promoter(DsbGp) 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

Related Parts

• ahpC(Part:BBa_K362001)
  • AhpCp1000(Part:BBa_K1104203)
    • Part:BBa_K1104243: AhpCp1000+E0840
  • AhpCp2D1(Part:BBa_K1104204)
    • Part:BBa_K1104244: AhpCp2D1+E0840
  • AhpCp2(Part:BBa_K1104205)
    • Part:BBa_K1104245: AhpCp2+E0840
  • AhpCpD1(Part:BBa_K1104206)
    • Part:BBa_K1104246: AhpCpD1+E0840
  • AhpCp1(Part:BBa_K1104207)
    • Part:BBa_K1104247: AhpCp1+E0840
  • DsbGp(Part:BBa_K1104208)

Sequence and Features