Difference between revisions of "Part:BBa K1104202"

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==Introduction==
 
==Introduction==
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HemHp promoter is controlled by OxyR (transcription factor), which is activated by ROS(Reactive Oxygen Species). An intramolecular disulfide bond between Cys199 and Cys208 will form in an OxyR transcription factor, and active OxyR molecule will bind to the binding site of HemHp, making the following circuit turned on.
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HemHp is a ROS-induced promoter controlled by OxyR (transcription factor) which is activated by ROS (Reactive Oxygen Species).  
  
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In our project: ''Bee. coli'', we use HemH as the sensor of ''Bee. coli'' to sense ''Nosema ceranae''. After ''Nosema ceranae'' enter bees' midgut, the natural immune reaction will lead to ROS released by epithelia of bees' midgut. We select several promoters that can be induced by OxyR, which is activated by ROS. One of these promoters is HemHp.
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An intramolecular disulfide bond between Cys199 and Cys208 will form in an OxyR transcription factor, and active OxyR molecule will bind to the binding site of HemHp, making the following circuit turned on. In our project: ''Bee. coli'', we use HemHp as the sensor of ''Bee. coli'' to sense ''Nosema ceranae''. After ''Nosema ceranae'' enter bees' midgut, the natural immune reaction will lead to ROS released by epithelia of bees' midgut. We select several promoters that can be induced by OxyR ([https://parts.igem.org/Part:BBa_K1104200 Part:BBa_K1104200]), which is activated by ROS. One of these promoters is HemHp.
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OxyR is an activator of ahpC promoter and its improvement parts. More details about OxyR can be found on the PartRegistry page: [https://parts.igem.org/Part:BBa_K1104200 Part:BBa_K1104200].
  
 
[[File: NYMU_HemH.png|frame|center|'''HemHp''']]
 
[[File: NYMU_HemH.png|frame|center|'''HemHp''']]
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HemHp is regulated by σ70, and there are two TFBSs upstream for OxyR.
 
HemHp is regulated by σ70, and there are two TFBSs upstream for OxyR.
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We also did some literature study, according to the citation.
 
  
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In ''E. coli'' genome, thioredoxin(trxC) will express in response to elevated H<sub>2</sub>O<sub>2</sub> level. Expression of genes encoding thioredoxin(trxC) is regulated by OxyR. Oxidized OxyR binds directly to HemHp and induce its expression.
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[[File: NYMU_ecoH.png|frame|center|Nucleotide Sequence of HemHp(taken from [http://ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-12951 Ecocyc])]]
  
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[[File: NYMU_phemh.png|frame|center|Schematic drawing of the HemH regulon in ''E. coli'' K12 (taken from [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-3104 Ecocyc])]][[File: NYMU_ecoH.png|frame|left|Nucleotide Sequence of HemHp(taken from [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-3104 Ecocyc])]]
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[[File: NYMU_ecoH1.png|frame|center|OxyR Sequence of HemHp(taken from [http://ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-12951 Ecocyc])]]
 
{|
 
{|
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|[[File: NYMU_ecoH1.png|frame|center|OxyR TFBS1 Sequence of HemHp(taken from [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-3104 Ecocyc])]]
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|[[File: NYMU_ecoH1.png|frame|center|OxyR TFBS1 Sequence of HemHp(taken from [http://ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-12951 Ecocyc])]]
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|[[File: NYMU_ecoH2.png|frame|center|OxyR TFBS2 Sequence of HemHp(taken from [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-3104 Ecocyc])]]
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|[[File: NYMU_ecoH2.png|frame|center|OxyR TFBS2 Sequence of HemHp(taken from [http://ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-12951 Ecocyc])]]
 
|}
 
|}
 
:::::<sub>Reference: Ritz, D. (January 28, 2000). Thioredoxin 2 Is Involved in the Oxidative Stress Response in Escherichia coli. Journal of Biological Chemistry, 275, 4, 2505-2512.</sub>
 
:::::<sub>Reference: Ritz, D. (January 28, 2000). Thioredoxin 2 Is Involved in the Oxidative Stress Response in Escherichia coli. Journal of Biological Chemistry, 275, 4, 2505-2512.</sub>
  
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==Usage in Our Project==
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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.
 
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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[[File: NYMU_HemHp-strenthening device.png|frame|center|'''HemHp-strenthening device''']]
 
[[File: NYMU_HemHp-strenthening device.png|frame|center|'''HemHp-strenthening device''']]
  
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==Related Parts==
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#[https://parts.igem.org/Part:BBa_K1104242 Part:BBa_K1104242]: HemHp+[https://parts.igem.org/Part:BBa_E0840 E0840], reporter with GFP for promoter testing.
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#[https://parts.igem.org/Part:BBa_K1104201 Part:BBa_K1104202]: HemHp, another OxyR-induced promoter.
  
 
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<!-- Add more about the biology of this part here
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===Usage and Biology===
 
  
 
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Latest revision as of 21:30, 4 October 2013

HemH promoter

Introduction

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

An intramolecular disulfide bond between Cys199 and Cys208 will form in an OxyR transcription factor, and active OxyR molecule will bind to the binding site of HemHp, making the following circuit turned on. In our project: Bee. coli, we use HemHp as the sensor of Bee. coli to sense Nosema ceranae. After Nosema ceranae enter bees' midgut, the natural immune reaction will lead to ROS released by epithelia of bees' midgut. We select several promoters that can be induced by OxyR (Part:BBa_K1104200), which is activated by ROS. One of these promoters is HemHp.

OxyR is an activator of ahpC promoter and its improvement parts. More details about OxyR can be found on the PartRegistry page: Part:BBa_K1104200.

HemHp

Information of HemHp on [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-3104 Ecocyc]

NYMU HemHpEcartoon.png

We get information of HemHp from [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-3104 Ecocyc],

HemHp is regulated by σ70, and there are two TFBSs upstream for OxyR.

Nucleotide Sequence of HemHp(taken from [http://ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-12951 Ecocyc])
OxyR Sequence of HemHp(taken from [http://ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-12951 Ecocyc])
OxyR TFBS1 Sequence of HemHp(taken from [http://ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-12951 Ecocyc])
OxyR TFBS2 Sequence of HemHp(taken from [http://ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-12951 Ecocyc])
Reference: Ritz, D. (January 28, 2000). Thioredoxin 2 Is Involved in the Oxidative Stress Response in Escherichia coli. Journal of Biological Chemistry, 275, 4, 2505-2512.

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.

Besides, in order to enhance the strength of HemHp, we added a device (more details on [http://2013.igem.org/Team:NYMU-Taipei/Project/Inhibition/Sensor Sensing Nosema] page).

HemHp-strenthening device

Related Parts

  1. Part:BBa_K1104242: HemHp+E0840, reporter with GFP for promoter testing.
  2. Part:BBa_K1104202: HemHp, another OxyR-induced promoter.

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]