Difference between revisions of "Part:BBa K1104205"
(→‎How ahpC (Part:BBa_K362001) is improved?)
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1104205 short</partinfo>
 
<partinfo>BBa_K1104205 short</partinfo>
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[[File: NYMU_AhpCp2.png|frame|center|'''AhpCp2''']]
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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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AhpCp2 is a ROS-induced promoter controlled by OxyR (transcription factor)which is activated by ROS (Reactive Oxygen Species).
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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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[[File: NYMU_Ax.png|thumb|600px|center|'''Improvement of ahpC promoter([https://parts.igem.org/Part:BBa_K362001 Part:K362001])''']]
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AhpCp2 is composed of AhpCp2 promoter and a dual-TFBS (Transcription Factor Binding Site) for OxyR binding.
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<!-- Add more about the biology of this part here
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[[File: NYMU_Ax.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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===How ahpC ([https://parts.igem.org/Part:BBa_K362001 Part:BBa_K362001]) is improved?===
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'''In this part, we succesfully seperated AhpCp2(TFBS included) out of ahpC promoter ([https://parts.igem.org/Part:BBa_K362001 Part:K362001]).'''
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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 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_K1104203 Part:BBa_K1104203]): 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.
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===Usage and Biology===
 
===Usage and Biology===
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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.
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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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[[File: NYMU_O12.png|frame|center|'''Strenthening device''']]
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===Related Parts===
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*ahpC([https://parts.igem.org/Part:BBa_K362001 Part:BBa_K362001)]
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**AhpCp1000([https://parts.igem.org/Part:BBa_K1104203 Part:BBa_K1104203])
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***[https://parts.igem.org/Part:BBa_K1104242 Part:BBa_K1104243]: AhpCp1000+[https://parts.igem.org/Part:BBa_E0840 E0840]
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**AhpCp2D1([https://parts.igem.org/Part:BBa_K1104204 Part:BBa_K1104204])
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***[https://parts.igem.org/Part:BBa_K1104244 Part:BBa_K1104244]: AhpCp2D1+[https://parts.igem.org/Part:BBa_E0840 E0840]
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**AhpCp2([https://parts.igem.org/Part:BBa_K1104205 Part:BBa_K1104205])
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***[https://parts.igem.org/Part:BBa_K1104245 Part:BBa_K1104245]: AhpCp2+[https://parts.igem.org/Part:BBa_E0840 E0840]
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**AhpCpD1([https://parts.igem.org/Part:BBa_K1104206 Part:BBa_K1104206])
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***[https://parts.igem.org/Part:BBa_K1104246 Part:BBa_K1104246]: AhpCpD1+[https://parts.igem.org/Part:BBa_E0840 E0840]
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**AhpCp1([https://parts.igem.org/Part:BBa_K1104207 Part:BBa_K1104207])
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***[https://parts.igem.org/Part:BBa_K1104247 Part:BBa_K1104247]: AhpCp1+[https://parts.igem.org/Part:BBa_E0840 E0840]
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**DsbGp([https://parts.igem.org/Part:BBa_K1104208 Part:BBa_K1104208])
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Latest revision as of 22:39, 4 October 2013

AhpCp2

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

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

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).

How ahpC (Part:BBa_K362001) is improved?

In this part, we succesfully seperated AhpCp2(TFBS included) out of ahpC promoter (Part:K362001).

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 extracting the AhpCp2(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.
  • 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




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]