Difference between revisions of "Part:BBa K1104208"

Revision as of 22:24, 4 October 2013

DsbGp

DsbGp

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

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

Improvement of ahpC promoter(Part:K362001)

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

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

How ahpC (Part:BBa_K362001) is improved?

In this part, we succesfully seperated DsbGp (TFBS included) out of ahpC promoter (Part:K362001).

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.
• AhpCp2 (Part:BBa_K1104205): Only one promoter(AhpCp2) 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