Device

Part:BBa_K1104250:Experience

Designed by: Ting-Yun Chiang   Group: iGEM13_NYMU-Taipei   (2013-10-27)

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Confirming expression of OxyR by BBa_K1104250

We conduct an SDS Polyacrylamide gel electrophoresis for OxyR expression.

NYMU SDS.png

Setting control group contains only E.coli MG1655 and experimental group with K1104250 added to AhpCp2D1(K1104204), the electrophoresis result shows OxyR (Molecular Weight of Polypeptide: 34.276 kD) can really be expressed.

Applications of BBa_K1104250

OxyR-included circuit(Device1+Device2)

In OxyR-included circuit, Device2 is composed of sensor (OxyR-induced promoter, including TrxCp, HemHp, sufA, AhpCp1000, AhpCp2D1, AhpCp2, AhpCpD1, AhpCp1, DsbGp) plus reporter (Part:BBa_E0840). In this case we use AhpCp2D1.

Device1 (Part:BBa_K1104250)
a constitutive promoter(Part:BBa_J23102)+ RBS(Part:BBa_B0034)+ activator OxyR(Part:BBa_K1104200)+ double terminator Part:BBa_B0015
Device2
a sensor (OxyR-induced promoter, including TrxCp, HemHp, sufA, AhpCp1000, AhpCp2D1, AhpCp2, AhpCpD1, AhpCp1, DsbGp) + 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), RBS(Part:BBa_B0034), activator OxyR(Part:BBa_K1104200),and double terminator Part:BBa_B0015.

Putting Device1 device:BBa_K1104250 upstream to promoter AhpCp2D1(Part:BBa_K1104204) remarkably enhanced the strength of promoter AhpCp2D1(Part:BBa_K1104204) as our experiment results shows:

NYMU 2D1O.png
NYMU 2D1Ovs.png


We designed circuit fighting against Nosema ceranae. To enhance the strength , we added a device (more details on Sensing Nosema page).

Strenthening device

Not only for sensing the midgut oxygen stress caused by Nosemasis, in our project, there are some previous iGEM team who used OxyR-induced promoter, such as 2010 KIT-Tokyo team using it for the ink of E.coli Pen colorful. And those OxyR-induced promoters relative to the projects.

Since ROS is a natural byproduct of the metabolism and have important roles in cell signaling and homeostasis, the improvement of ROS-induced promoter is useful for sensing some specific conditions. ROS is found to involve in cancer, aging and apoptosis. Thus ROS-induced promoter in the future application can be apply to diagnosis and therapy of certain kinds of physical phenomena or disease.

Devasagaya, TPA; Tilak JC, Boloor KK, Sane Ketaki S, Ghaskadbi Saroj S, Lele RD (October 2004). "Free Radicals and Antioxidants in Human Health: Current Status and Future Prospects". Journal of Association of Physicians of India (JAPI) 52: 796.

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