Difference between revisions of "Part:BBa K3733045"

 
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Use a constitutive promoter (BBa_J23110), RNA thermometer (BBa_K3733011), strong RBS (BBa_B0034), fluorescent protein neGFP (BBa_K3733012),and an artificial terminator (BBa_B1006). Will express high levels of fluorescent protein below 28 ℃.
 
Use a constitutive promoter (BBa_J23110), RNA thermometer (BBa_K3733011), strong RBS (BBa_B0034), fluorescent protein neGFP (BBa_K3733012),and an artificial terminator (BBa_B1006). Will express high levels of fluorescent protein below 28 ℃.
  
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===Usage and Biology===
 
===Usage and Biology===
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This is a composite part constructed to verify the effectiveness of the heat-repressible RNA thermometer.
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</p>
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===Functional Parameters===
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<p>
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We transformed this composite part into <i>E.coli</i> DH5α. We cultured both experimental group and control group at both 37 ℃ and 28 ℃ for 12 hours and detected RFU / OD<sub>600</sub> by a microplate reader. As the result shown in <b>Figure 1</b>, RFU / OD<sub>600</sub> data of the media at 28 ℃ are higher than ones at 37 ℃, which could explain the RNA thermometer is valid.
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<meta charset="utf-8">
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<title>无标题文档</title>
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<center><img src="https://static.igem.org/mediawiki/parts/c/c4/T--HZAU-China-HepT-comp-3.png" style="width:500px;height:360px"></center>
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<center><b>Figure 1. </b> RFU / OD<sub>600</sub> data of the media at 37 ℃ and 28 ℃(heat-repressible RNA thermosensors) </center>
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===Sequence and Features===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3733045 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3733045 SequenceAndFeatures</partinfo>
  

Revision as of 17:46, 20 October 2021


RNA Thermometer(heat-repressible) test system

Use a constitutive promoter (BBa_J23110), RNA thermometer (BBa_K3733011), strong RBS (BBa_B0034), fluorescent protein neGFP (BBa_K3733012),and an artificial terminator (BBa_B1006). Will express high levels of fluorescent protein below 28 ℃.

Usage and Biology

This is a composite part constructed to verify the effectiveness of the heat-repressible RNA thermometer.

Functional Parameters

We transformed this composite part into E.coli DH5α. We cultured both experimental group and control group at both 37 ℃ and 28 ℃ for 12 hours and detected RFU / OD600 by a microplate reader. As the result shown in Figure 1, RFU / OD600 data of the media at 28 ℃ are higher than ones at 37 ℃, which could explain the RNA thermometer is valid.

无标题文档

Figure 1. RFU / OD600 data of the media at 37 ℃ and 28 ℃(heat-repressible RNA thermosensors)

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 149
    Illegal AgeI site found at 272
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 82