Difference between revisions of "Part:BBa K1104204:Experience"

Revision as of 10:07, 27 October 2013

This part: AhpCp2D1 is one of the improvement versions of BioBrick Part: ahpC promoter (Part:K362001) designed by [http://2010.igem.org/Team:KIT-Kyoto/Parts 2010 KIT-Tokyo team]. AhpCp2D1 (Part:BBa_K1104204) was used as sensor to ROS (Reactive Oxygen Species). More details about ROS-induced promoters and part improvements of ahpC promoter are in page:[http://2013.igem.org/Team:NYMU-Taipei/Project/Inhibition/Sensor Sensing Nosema ceranae].

AhpCp2D1 +E0840 (Part:BBa_K1104244)

We use the device: AhpCp2D1 +E0840 (Part:BBa_K1104244) includes promoter (Part:BBa_K1104204) and Part:BBa_E0840, induce the promoter by different concentrations of H₂O₂, and identify the strength. After AhpCp1000 (Part:BBa_K1104204) is activated, GFP generator BBa_E0840 takes as input a transcriptional signal (PoPS) and produce as output the fluorescent protein GFP.

We tested the improved promoter: AhpCp2D1 by following these [http://2013.igem.org/wiki/index.php?title=Team:NYMU-Taipei/Experiments/Protocols&oldid=243384 Promoter Testing protocols]. This wiki page shows the protocols we used for promoter testing.

Results

This is the relative promoter strength of BBa_K1104204 under different H₂0₂ concentrations tested using BBa_K1104244 (BBa_K1104204 +E0840) and found that there is a trend where the higher the H₂0₂ concentration, the higher the promoter expression:

Strengths of the basal level of various promoters that respond to reactive oxygen species with respect to BBa_J23101, and the AhpC2D1 promoter shows higher strength than the AhpCp 1000 which is an effective improvement :

Comparison between ahpC and its improvements. (Part:BBa_K1104244)

Promoter testing of ahpC promoter (Part:K362001) and its improvements.

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: [Image:NYMU_2D1O.png|frame|center]]

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)

Discussion

Applications of BBa_K1104204

User Reviews

UNIQ1ca584cf9fc68d33-partinfo-00000000-QINU UNIQ1ca584cf9fc68d33-partinfo-00000001-QINU

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 Promoter testing of ahpC promoter ([https://parts.igem.org/Part:BBa_K362001 Part:K362001]) and its improvements.
 [[File: NYMU_AA.png|frame|center]]
+Device1 in order to enhance the effect of ROS on ''E. coli'' is added ahead: 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]),and double terminator [https://parts.igem.org/Part:BBa_B0015 Part:BBa_B0015]. Putting Device1 [https://parts.igem.org/Part:BBa_K1104250 device: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: [Image:NYMU_2D1O.png|frame|center]]
+{|
+|[[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])]]
+|}
 ==Discussion==