Difference between revisions of "Part:BBa K1104204"

Revision as of 20:44, 4 October 2013

AhpCp2D1

AhpCp2D1 is a ROS-induced promoter, which is controlled by OxyR (transcription factor) activated by ROS (Reactive Oxygen Species).

Mutation of ahpC promoter(Part:K362001)

AhpCp2D1 is composed of 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 and DsbGp.

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 AhpCp2D1, we succesfully mutated the PstI cutting site (ctgcag->ctacag) of ahpC promoter (Part:K362001), and removed the dsbG coding sequence.

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.

Where is AhpCp2D1 improved?

In this part,the PstI cutting site in ahpCp (BBa_K362001) is mutated at one point.

How ahpCp (Part:BBa_K362001) is improved?

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

AhpCp1000 (Part:BBa_K1104204): The PstI cutting site is mutated.
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 are left.
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 4: / Line 4: @@
 AhpCp2D1 is a ROS-induced promoter, which is controlled by OxyR (transcription factor) activated by ROS (Reactive Oxygen Species).
 [[File: NYMU_A2D1x.png|thumb|600px|center|'''Mutation of ahpC promoter([https://parts.igem.org/Part:BBa_K362001 Part:K362001])''']]
-AhpCp2D1 is composed of 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]). There are also two dual-TFBSs (Transcription Factor Binding Site) for OxyR binding between AhpCp2 ([http://ptruncated coding sequence from the promoter sequencetruncated coding sequence from the promoter sequencetruncated coding sequence from the promoter sequencebG coding)from the promoter sequence.
+AhpCp2D1 is composed of 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]). There are also two dual-TFBSs (Transcription Factor Binding Site) for OxyR binding between AhpCp2 and DsbGp.
 ==Improvement==
@@ Line 12: / Line 12: @@
 ahpC promoter, as well as its improvement, can be activated by OxyR ([https://parts.igem.org/Part:BBa_K1104200 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 ([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 dsbG coding removed.
+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 ([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 dsbG coding removed.
 ===Where is AhpCp2D1 improved?===
 In this part,the PstI cutting site in ahpCp (BBa_K362001) is mutated at one point.
@@ Line 20: / Line 21: @@
 *AhpCp1000 ([https://parts.igem.org/Part:BBa_K1104204 Part:BBa_K1104204]): The PstI cutting site is mutated.
 *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.
+*AhpCpD1 ([https://parts.igem.org/Part:BBa_K1104206 Part:BBa_K1104206]): Bidirectional promoter: AhpCp1 and DsbGp(reverse promoter), and their shared TFBS are left.
 *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.