Difference between revisions of "Part:BBa K1104200:Experience"

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.

Device1(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.

Applications of BBa_K1104200

OxyR-included circuit(Device1+Device2)

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).

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 [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.

User Reviews

UNIQ612ce9595b8cdae4-partinfo-00000000-QINU UNIQ612ce9595b8cdae4-partinfo-00000001-QINU

@@ Line 5: / Line 5: @@
 We conduct an SDS Polyacrylamide gel electrophoresis for OxyR expression.
 [[File: NYMU_SDS.png|frame|center]]
+[[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.
 ===Applications of BBa_K1104200===
 [[Image:NYMU_O12.png|frame|center|'''OxyR-included circuit(Device1+Device2)''']]
@@ Line 19: / Line 21: @@
 {|
-|[[Image:NYMU_O1.png|frame|center|'''Device1'''<br>a constitutive promoter([https://parts.igem.org/Part:BBa_J23102 Part:BBa_J23102])+ RBS([https://parts.igem.org/Part:BBa_B0034 Part:BBa_B0034])+ activator OxyR([https://parts.igem.org/Part:BBa_K1104200 Part:BBa_K1104200])+ double terminator [https://parts.igem.org/Part:BBa_B0015 Part:BBa_B0015]]]
+|[[Image:NYMU_O1.png|frame|center|'''Device1([https://parts.igem.org/Part:BBa_K1104250 Part:BBa_K1104250])'''<br>a constitutive promoter([https://parts.igem.org/Part:BBa_J23102 Part:BBa_J23102])+ RBS([https://parts.igem.org/Part:BBa_B0034 Part:BBa_B0034])+ activator OxyR([https://parts.igem.org/Part:BBa_K1104200 Part:BBa_K1104200])+ double terminator [https://parts.igem.org/Part:BBa_B0015 Part:BBa_B0015]]]
 |[[Image:NYMU_O2.png|frame|center|'''Device2'''<br>a sensor (OxyR-induced promoter, including TrxCp, HemHp, sufA, AhpCp1000, AhpCp2D1, AhpCp2, AhpCpD1, AhpCp1, DsbGp) + reporter ([https://parts.igem.org/Part:BBa_E0840 Part:BBa_E0840])]]
 |}
 Putting Device1 [https://parts.igem.org/Part:BBa_K1104250 BBa_K1104250] upstream to promoter AhpCp2D1([https://parts.igem.org/Part:BBa_K1104204 Part:BBa_K1104204]) remarkably enhanced the strength of promoter AhpCp2D1([https://parts.igem.org/Part:BBa_K1104204 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).
+[[File: NYMU_O12.png|thumb|500px|center|'''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 [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.
 ===User Reviews===