Regulatory

Part:BBa_K1104207

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

AhpCp1

Mutation of ahpC promoter(Part:K362001)

AhpCp1 is a ROS-induced promoter, which is controlled by OxyR (transcription factor) activated by ROS (Reactive Oxygen Species).

AhpCp1 is composed of AhpCp1 promoter and a dual-TFBS (Transcription Factor Binding Site) for OxyR binding.

Improvement of ahpC promoter(Part:K362001)

OxyR is activator of AhpCp1 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 2010 KIT-Tokyo team. On PartRegistry, the complex part(according to Ecocyc) composition contains hybrid promoters, shared TFBS (Transcription Factor Binding Site), and reverse promoter DsbG. In this part AhpCp1, we succesfully seperated AhpCp1(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 AhpCp1 (TFBS included) out of ahpC promoter (Part:K362001).

We annotated it thouroughly based on data from (Ecocyc), and found that it contains dsbG coding sequence, AhpCp1 (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 AhpCp1(TFBS included).

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.
  • AhpCpD1 (Part:BBa_K1104206): Bidirectional promoter: AhpCp1 and DsbGp(reverse promoter), and their shared TFBS.
  • AhpCp2 (Part:BBa_K1104207): Only one promoter(AhpCp2) 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 Killing Nosema page) in the following circuit will express.

To enhance the strength , we added a device (more details on Sensing Nosema page).

Strenthening device

Related Parts


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