Difference between revisions of "Part:BBa K1104203"

Revision as of 20:01, 4 October 2013

AhpCp1000

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

AhpCp1000 is composed 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 Pst1 site (ctgcag->ctacag) of ahpC promoter (Part:K362001).

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

Where is AhpCp1000 improved?

The PstI cutting site in ahpCp (BBa_K362001)

How ahpCp (Part:BBa_K362001) is improved?

We annotated it thouroughly based on data from ([http://ecocyc.org/ECOLI/new-image?object=EG11384 Ecocyc 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 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)
  - Part:BBa_K1104248: DsbGp+E0840

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]

@@ Line 15: / Line 15: @@
 ahpC promoter, as well as its improvement, can be activated by OxyR ([https://parts.igem.org/Part:BBa_K1104200 Part:BBa_K1104200]).
+===Where is AhpCp1000 improved?===
+The PstI cutting site in ahpCp (BBa_K362001)
+===How ahpCp ([https://parts.igem.org/Part:BBa_K362001 Part:BBa_K362001]) is improved?===
+We annotated it thouroughly based on data from ([http://ecocyc.org/ECOLI/new-image?object=EG11384 Ecocyc Ecocyc]), and found that it contains dsbG coding sequence, AhpCp2 ([https://parts.igem.org/Part:BBa_K1104205 Part:BBa_K1104205]), reverse promoter DsbGp ([https://parts.igem.org/Part:BBa_K1104208 Part:BBa_K1104208]),and AhpCp1 ([https://parts.igem.org/Part:BBa_K1104207 Part:BBa_K1104207]), and a PstI cutting site. Thus we improved the promoter by first mutating the PstI cutting site in ahpCp  ([https://parts.igem.org/Part:BBa_K362001 Part:BBa_K362001]) and make AhpCp1000.
+Here is the overview about the other ahpC promoter ([https://parts.igem.org/Part:BBa_K362001 Part:BBa_K362001]) improvements:
+*AhpCp2D1 ([https://parts.igem.org/Part:BBa_K1104204 Part:BBa_K1104204]): After mutating the PstI cutting site, the truncated coding sequence from the promoter sequence is removed.
+*AhpCp2 ([https://parts.igem.org/Part:BBa_K1104205 Part:BBa_K1104205]): Only one promoter(AhpCp2) and its TFBS.
+*AhpCpD1 ([https://parts.igem.org/Part:BBa_K1104206 Part:BBa_K1104206]): Bidirectional promoter: AhpCp1 and DsbGp(reverse promoter), and their shared TFBS.
+*AhpCp1 ([https://parts.igem.org/Part:BBa_K1104207 Part:BBa_K1104207]): Only one promoter(AhpCp1) and its TFBS.
+*DsbGp ([https://parts.igem.org/Part:BBa_K1104208 Part:BBa_K1104208]): Only the reverse promoter(DsbGp) and its TFBS.
 ===Usage and Biology===
@@ Line 23: / Line 35: @@
 ===Related Parts===
 *ahpC([https://parts.igem.org/Part:BBa_K362001 Part:BBa_K362001)]
 **AhpCp1000([https://parts.igem.org/Part:BBa_K1104203 Part:BBa_K1104203])