Regulatory

Part:BBa_K1104201

Designed by: Ting-Yun Chiang   Group: iGEM13_NYMU-Taipei   (2013-09-11)
Revision as of 16:14, 30 September 2013 by Windy6632 (Talk | contribs) (→‎Introduction)

TrxC promoter

Introduction

TrxC 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 TrxCp, making the following circuit turned on.

In our project: Bee. coli, we use TrxCp 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 TrxCp.

TrxCp

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

NYMU TrxCpEcartoon.png

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

TrxCp is regulated by σ70, and there are two TFBSs upstream for OxyR. We also did some literature study, according to the citation.

In E. coli genome, thioredoxin(trxC) will express in response to elevated H2O2 level. Expression of genes encoding thioredoxin(trxC) is regulated by OxyR. Oxidized OxyR binds directly to TrxCp and induce its expression.

Schematic drawing of the TrxC regulon in E. coli K12 (taken from [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-3104 Ecocyc])
Nucleotide Sequence of TrxCp(taken from [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-3104 Ecocyc])
OxyR TFBS1 Sequence of TrxCp(taken from [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-3104 Ecocyc])
OxyR TFBS2 Sequence of TrxCp(taken from [http://www.ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU0-3104 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 in Our Project

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 TrxCp, we added a device (more details on [http://2013.igem.org/Team:NYMU-Taipei/Project/Inhibition/Sensor Sensing Nosema] page).

TrxCp-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]


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Parameters
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