Composite

Part:BBa_K3733045

Designed by: Zhenhao Han   Group: iGEM21_HZAU-China   (2021-10-01)


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.

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

Sequence and Features

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


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Categories
Parameters
None