Part:BBa_K1104200

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. The OxyR gene, a positive regulatory gene for the oxidative response, then controls proteins which are induced by treatment with H₂O₂.

OxyR(BBa_K1104200)

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.

• ahpC(Part:BBa_K362001) is improved its complex composition and Assembly Compatibility by us. We designed ahpC(Part:BBa_K362001) improvement into 6 versions:
  • AhpCp1000(Part:BBa_K1104203)
  • AhpCp2D1(Part:BBa_K1104204)
  • AhpCp2(Part:BBa_K1104205)
  • AhpCpD1(Part:BBa_K1104206)
  • AhpCp1(Part:BBa_K1104207)
  • DsbGp(Part:BBa_K1104208)

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)

• • AhpCp1000(Part:BBa_K1104203)
    • Part:BBa_K1104243: AhpCp1000+E0840
  • AhpCp2D1(Part:BBa_K1104204)
    • Part:BBa_K1104244: AhpCp2D1+E0840
  • AhpCp2(Part:BBa_K1104205)
    • Part:BBa_K1104245: AhpCp2+E0840
  • AhpCpD1(Part:BBa_K1104206)
    • Part:BBa_K1104246: AhpCpD1+E0840
  • AhpCp1(Part:BBa_K1104207)
    • Part:BBa_K1104247: AhpCp1+E0840
  • DsbGp(Part:BBa_K1104208)
    • Part:BBa_K1104248: DsbGp+E0840

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]