Part:BBa_K3733046

RNA Thermometer(heat-inducible) test system

Use a constitutive promoter (BBa_J23110), RNA thermometer (BBa_K3247005), strong RBS (BBa_B0034), fluorescent protein neGFP (BBa_K3733012),and an artificial terminator (BBa_B1006). Will express high levels of fluorescent protein above 37 ℃. Source:Used a strong RBS to produce high levels of expression of the neGFP fluorescent protein.

Usage and Biology

This is a composite part constructed to verify the effectiveness of the heat-inducible 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 37 ℃ are obviously higher than ones at 28 ℃, which could explain the RNA thermometer is valid.

Figure 1. RFU / OD₆₀₀ data of the media at 37 ℃ and 28 ℃(heat-inducible 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 158
  Illegal AgeI site found at 281
  
• 1000
  COMPATIBLE WITH RFC[1000]