Coding

Part:BBa_K1104200

Designed by: Ting-Yun Chiang   Group: iGEM13_NYMU-Taipei   (2013-09-11)
Revision as of 18:23, 1 October 2013 by Windy6632 (Talk | contribs) (Usage and Biology)

OxyR

Introduction

OxyR is Escherichia coli transcription factor that senses H2O2 to activate transcription, and is activated through the formation of an intramolecular disulfide bond.

Literature Study of OxyR

Illustration of OxyR Activation

OxyR, a positive regulator of hydrogen peroxide-inducible genes in E. coli. The OxyR transcription factor regulates the H2O2 response of E. coli. H2O2 oxidizes OxyR Cys199 to an R-SOH that reacts with Cys208 in an intramolecular disulphide bond.

Structure of the OxyR Regulatory Domain
(A)Reduced Form: Inactive
(B)Oxidized Form: Activated by ROS
Reference: D'Autréaux, B., & Toledano, M. B. (October 01, 2007). ROS as signalling molecules: mechanisms that generate specificity in ROS homeostasis. Nature Reviews Molecular Cell Biology, 8, 10, 813-824.


Usage and Biology

After Nosema ceranae enter bees' midgut, the natural immune reaction will lead to ROS released by epithelia of bees' midgut. In our project: Bee. coli, we use OxyR-induced promoter as the sensor of Bee. coli to sense Nosema ceranae. We select several promoters that can be induced by OxyR(Part:BBa_K1104200).

There are promoters that can be activated by OxyR: TrxCp(Part:BBa_K1104201), HemHp(Part:BBa_K1104202), ahpC(Part:BBa_K362001), sufA(Part:BBa_K362005), and dps(Part:BBa_K362002), yiaA(Part:BBa_K362006)

The OxyR-induced promoters:

  • TrxCp(Part:BBa_K1104201) and HemHp(Part:BBa_K1104202) are parts created by our team [http://2013.igem.org/Team:NYMU-Taipei/Modeling/MainParts NYMU_Taipei].(Come to the [http://2013.igem.org/Team:NYMU-Taipei/Experiment/Wet_Lab Wet Lab] page to see our part list.)
  • sufA(Part:BBa_K362005), designed by [http://2010.igem.org/Team:KIT-Kyoto/Parts 2010 KIT-Tokyo team], is successfully characterized again by us.

In order to enhance the strength and sensitivity of ROS-sensing promoters, we make E. coli continuously produce inactivated OxyR transcription factors, so that ROS can affect promoters more easily.

We designed a device that is composed of a sensor (OxyR-induced promoter, including TrxCp, HemHp, sufA, AhpCp1000, AhpCp2D1, AhpCp2, AhpCpD1, AhpCp1, DsbGp) plus reporter (Part:BBa_E0840). Device1 in order to enhance the effect of ROS on E. coli is added ahead: a constitutive promoter(Part:BBa_J23102), and OxyR(Part:BBa_K1104200).

Related Parts

  1. TrxCp(Part:BBa_K1104201)
  2. Part:BBa_K1104241: TrxCp+E0840
  3. HemHp(Part:BBa_K1104202)
  4. Part:BBa_K1104242: HemHp+E0840
  5. sufA(Part:BBa_K362005)
  6. ahpC(Part:BBa_K362001)

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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Categories
Parameters
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