Difference between revisions of "Part:BBa K3431050"
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<partinfo>BBa_K3431050 short</partinfo>
 
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The is a sfGFP (BBa_K3431031) generating device regulated by Banana Toehold Switch (BBa_K3431039), with T7 Promoter (BBa_I719005) and T7 Terminator (BBa_K731721) as recognition sites for T7 RNA Polymerase in PURExpress in vitro protein synthesis kit.
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=== Introduction ===
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The sequence of this part is from a modular synthetic biology education kit, banana toehold sensor sfGFP (Biobits) 1 . This composite part can be triggered by the RNA of banana and express sfGFP (BBa_K3431031) as its reporter.
  
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===Experiment result===
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===Characterization===
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The 2020 iGEM CSMU_Taiwan carried out a functionality test on this part, measuring the real-time fluorescence of the sfGFP within the 80 minutes protein production process. The result is shown below.
 
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<img src="https://static.igem.org/mediawiki/parts/f/fc/T--CSMU_Taiwan--Fig._2_%28EXP_1%29.png" style="width:100%">
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<img src="https://static.igem.org/mediawiki/parts/f/fc/T--CSMU_Taiwan--Fig._2_%28EXP_1%29.png" style="width:40%">
 
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Figure 1. The relative fluorescence of the sfGFP in the 80 minutes protein production process. The relative fluorescence was measured by the Synergy H1 Hybrid Multi-Mode Reader. The green line refers to the conditions with banana triggers, while the blue line refers to the conditions without a banana trigger.
 
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<b>Results</b> As seen, the sfGFPs would be highly expressed when the banana triggers existed, while they would be inhibited when there was no trigger in the environment. This suggested the regulatory function of the toehold switch.
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===Reference===
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1. Huang, A., Nguyen, P. Q., Stark, J. C., Takahashi, M. K., Donghia, N.,Ferrante, T., Dy, A. J., Hsu, K. J., Dubner, R. S., Pardee, K., Jewett, M. C., &amp;Collins, J. J. (2018). BioBits™ Explorer: A modular synthetic biology education kit. Science advances, 4(8), eaat5105. https://doi.org/10.1126/sciadv.aat5105
  
 
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Revision as of 09:05, 25 October 2020


Banana_ToeholdSwitch-Regulated sfGFP

Introduction

The sequence of this part is from a modular synthetic biology education kit, banana toehold sensor sfGFP (Biobits) 1 . This composite part can be triggered by the RNA of banana and express sfGFP (BBa_K3431031) as its reporter.

Characterization

The 2020 iGEM CSMU_Taiwan carried out a functionality test on this part, measuring the real-time fluorescence of the sfGFP within the 80 minutes protein production process. The result is shown below.

Figure 1. The relative fluorescence of the sfGFP in the 80 minutes protein production process. The relative fluorescence was measured by the Synergy H1 Hybrid Multi-Mode Reader. The green line refers to the conditions with banana triggers, while the blue line refers to the conditions without a banana trigger.
Results As seen, the sfGFPs would be highly expressed when the banana triggers existed, while they would be inhibited when there was no trigger in the environment. This suggested the regulatory function of the toehold switch.

Reference

1. Huang, A., Nguyen, P. Q., Stark, J. C., Takahashi, M. K., Donghia, N.,Ferrante, T., Dy, A. J., Hsu, K. J., Dubner, R. S., Pardee, K., Jewett, M. C., &Collins, J. J. (2018). BioBits™ Explorer: A modular synthetic biology education kit. Science advances, 4(8), eaat5105. https://doi.org/10.1126/sciadv.aat5105

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 865
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]