Difference between revisions of "Part:BBa K1104205"

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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1104205 short</partinfo>
 
<partinfo>BBa_K1104205 short</partinfo>
[[File: NYMU_AhpCp2.png|frame|center|'''AhpCp2''']]
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AhpCp2 is a ROS-induced promoter, which is controlled by OxyR (transcription factor) activated by ROS (Reactive Oxygen Species).
We improve the function of a BioBrick Part: AhpC promoter([https://parts.igem.org/Part:BBa_K362001 K362001])designed by [http://2010.igem.org/Team:KIT-Kyoto/Parts 2010 KIT-Tokyo team] into four versions.On PartRegistry, the complex part(according to [http://ecocyc.org/ECOLI/new-image?object=EG11384 Ecocyc]) composition contains hybrid promoters(AhpC1, AhpC2, and DsbG promoter), shared TFBS (Transcription Factor Binding Site), and reverse promoter DsbG.
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There is only AhpC2 promoter in this part. We designed the new part and test the promoter separately.
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AhpCp2 is composed of AhpCp2 promoter and a dual-TFBS (Transcription Factor Binding Site) for OxyR binding.
[[File: NYMU_Ax.png|thumb|600px|center|'''Improvement of ahpC promoter([https://parts.igem.org/Part:BBa_K362001 Part:K362001])''']]
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[[File: NYMU_A2x.png|thumb|600px|center|'''Mutation of ahpC promoter([https://parts.igem.org/Part:BBa_K362001 Part:K362001])''']]
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OxyR is activator of AhpCp2 promoter. More details about OxyR can be found on the PartRegistry page: [https://parts.igem.org/Part:BBa_K1104200 Part:BBa_K1104200].
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==Improvement==
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 +
We improved a BioBrick Part: ahpC promoter ([https://parts.igem.org/Part:BBa_K362001 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 AhpCp2, we succesfully seperated AhpCp2(TFBS included) out of ahpC promoter ([https://parts.igem.org/Part:BBa_K362001 Part:K362001]).
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ahpC promoter, as well as its improvement, can be activated by OxyR ([https://parts.igem.org/Part:BBa_K1104200 Part:BBa_K1104200]).
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===Where is AhpCp1000 improved?===
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In this part,the PstI cutting site in ahpCp (BBa_K362001) is mutated at one point.
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===How ahpCp ([https://parts.igem.org/Part:BBa_K362001 Part:BBa_K362001]) is improved?===
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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 extracting the AhpCp2(TFBS included).  
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Here is the overview about the other ahpC promoter ([https://parts.igem.org/Part:BBa_K362001 Part:BBa_K362001]) improvements:
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*AhpCp1000 ([https://parts.igem.org/Part:BBa_K1104204 Part:BBa_K1104204]): The PstI cutting site is mutated.
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*AhpCp2D1 ([https://parts.igem.org/Part:BBa_K1104204 Part:BBa_K1104204]): After mutating the PstI cutting site, the truncated coding sequence from the DsbG promoter sequence is removed.
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*AhpCpD1 ([https://parts.igem.org/Part:BBa_K1104206 Part:BBa_K1104206]): Bidirectional promoter: AhpCp1 and DsbGp(reverse promoter), and their shared TFBS.
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*AhpCp1 ([https://parts.igem.org/Part:BBa_K1104207 Part:BBa_K1104207]): Only one promoter(AhpCp1) and its TFBS.
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*DsbGp ([https://parts.igem.org/Part:BBa_K1104208 Part:BBa_K1104208]): Only the reverse promoter(DsbGp) and its TFBS.
  
 
===Usage and Biology===
 
===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 [https://parts.igem.org/Part:BBa_K1104300 Defensin(Part:BBa_K1104300)], [https://parts.igem.org/Part:BBa_K1104301  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.
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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 [https://parts.igem.org/Part:BBa_K1104300 Defensin (Part:BBa_K1104300)], [https://parts.igem.org/Part:BBa_K1104301  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).
 
To enhance the strength , we added a device (more details on [http://2013.igem.org/Team:NYMU-Taipei/Project/Inhibition/Sensor Sensing Nosema] page).
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===Related Parts===
 
===Related Parts===
 
 
*ahpC([https://parts.igem.org/Part:BBa_K362001 Part:BBa_K362001)]
 
*ahpC([https://parts.igem.org/Part:BBa_K362001 Part:BBa_K362001)]
 
**AhpCp1000([https://parts.igem.org/Part:BBa_K1104203 Part:BBa_K1104203])
 
**AhpCp1000([https://parts.igem.org/Part:BBa_K1104203 Part:BBa_K1104203])
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**DsbGp([https://parts.igem.org/Part:BBa_K1104208 Part:BBa_K1104208])
 
**DsbGp([https://parts.igem.org/Part:BBa_K1104208 Part:BBa_K1104208])
 
***[https://parts.igem.org/Part:BBa_K1104248 Part:BBa_K1104248]: DsbGp+[https://parts.igem.org/Part:BBa_E0840 E0840]
 
***[https://parts.igem.org/Part:BBa_K1104248 Part:BBa_K1104248]: DsbGp+[https://parts.igem.org/Part:BBa_E0840 E0840]
<!-- Add more about the biology of this part here
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 +
===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 [https://parts.igem.org/Part:BBa_K1104300 Defensin(Part:BBa_K1104300)], [https://parts.igem.org/Part:BBa_K1104301  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).
 +
[[File: NYMU_O12.png|frame|center|'''Strenthening device''']]
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<!-- -->
 
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Revision as of 21:01, 4 October 2013

AhpCp2 AhpCp2 is a ROS-induced promoter, which is controlled by OxyR (transcription factor) activated by ROS (Reactive Oxygen Species).

AhpCp2 is composed of AhpCp2 promoter and a dual-TFBS (Transcription Factor Binding Site) for OxyR binding.

File:NYMU A2x.png
Mutation of ahpC promoter(Part:K362001)

OxyR is activator of AhpCp2 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 AhpCp2, we succesfully seperated AhpCp2(TFBS included) out of ahpC promoter (Part:K362001).

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

Where is AhpCp1000 improved?

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

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 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.
  • 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

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


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]