Part:BBa_K1104206

AhpCpD1

AhpCpD1

AhpCpD1 is a ROS-induced promoter controlled by OxyR (transcription factor)which is activated by ROS (Reactive Oxygen Species).

Improvement of ahpC promoter(Part:K362001)

AhpCpD1 is composed of reverse promoter DsbGp (Part:BBa_K1104208)and AhpCp1 (Part:BBa_K1104207). There are also a dual-TFBS (Transcription Factor Binding Site) for OxyR binding between DsbGp and AhpCp1.

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 AhpCpD1, we succesfully mutated the PstI cutting site (ctgcag->ctacag) of ahpC promoter (Part:K362001), and removed the dsbG coding sequence, then removed the AhpCp2(TFBS included).

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, then removed the AhpCp2(TFBS included).

How ahpCp (Part:BBa_K362001) is improved?

In this part, AhpCp1 and DsbGp are left, making this part a bidirectional hybrid promoter which has the ability to transcription in both directions.

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

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]