Difference between revisions of "Part:BBa K1104205"

Revision as of 21:04, 4 October 2013

AhpCp2 AhpCp2 is a ROS-induced promoter, which is controlled by OxyR (transcription factor) activated by ROS (Reactive Oxygen Species).

AhpCp2 is composed of AhpCp2 promoter and a dual-TFBS (Transcription Factor Binding Site) for OxyR binding.

Mutation of ahpC promoter(Part:K362001)

OxyR is activator of AhpCp2 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 AhpCp2, we succesfully seperated AhpCp2(TFBS included) out of ahpC promoter (Part:K362001).

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

Where is AhpCp1000 improved?

In this part,the PstI cutting site in ahpCp (BBa_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 extracting the AhpCp2(TFBS included).

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

• AhpCp1000 (Part:BBa_K1104204): 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.
• 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)
    • Part:BBa_K1104248: DsbGp+E0840

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

Sequence and Features