Regulatory

Part:BBa_K1104202

Designed by: Ting-Yun Chiang   Group: iGEM13_NYMU-Taipei   (2013-09-11)

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]


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Categories
Parameters
None