Difference between revisions of "Part:BBa K1104200:Experience"
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We conduct an SDS Polyacrylamide gel electrophoresis for OxyR expression. | We conduct an SDS Polyacrylamide gel electrophoresis for OxyR expression. | ||
[[File: NYMU_SDS.png|frame|center]] | [[File: NYMU_SDS.png|frame|center]] | ||
| − | [[Image:NYMU_O1.png|frame|center|'''Device1([https://parts.igem.org/Part:BBa_K1104250 Part:BBa_K1104250])''' | + | [[Image:NYMU_O1.png|frame|center|'''Device1([https://parts.igem.org/Part:BBa_K1104250 Part:BBa_K1104250])''']] |
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. | 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_K1104200=== | ===Applications of BBa_K1104200=== | ||
| + | |||
[[Image:NYMU_O12.png|frame|center|'''OxyR-included circuit(Device1+Device2)''']] | [[Image:NYMU_O12.png|frame|center|'''OxyR-included circuit(Device1+Device2)''']] | ||
Latest revision as of 17:49, 27 October 2013
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SDS-PAGE
We conduct an SDS Polyacrylamide gel electrophoresis for OxyR expression.
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_K1104200
In order to enhance the strength and sensitivity of ROS-sensing promoters, we make E. coli continuously produce inactivated OxyR, so that ROS can affect promoters more easily by adding a device: Device1(Part:BBa_K1104250).
Device1 in order to enhance the effect of ROS on E. coli is composed of :
- a constitutive promoter(Part:BBa_J23102)
- RBS(Part:BBa_B0034)
- activator OxyR(Part:BBa_K1104200)
- double terminator [https://parts.igem.org/Part:BBa_B0015 Part:BBa_B0015
| 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) |
Putting Device1 BBa_K1104250 upstream to promoter AhpCp2D1(Part:BBa_K1104204) remarkably enhanced the strength of promoter AhpCp2D1(Part:BBa_K1104204) as our experiment results shows.
We designed circuit fighting against Nosema ceranae. To enhance the strength , we added a device (more details on [http://2013.igem.org/Team:NYMU-Taipei/Project/Inhibition/Sensor Sensing Nosema] page).
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 [http://2010.igem.org/Team:KIT-Kyoto/Parts 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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