Difference between revisions of "Part:BBa K115003"
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===Characterization===
 
===Characterization===
 
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<p>To characterize this RNA thermometer in vivo, we made an RNA thermometer testing circuit, transformed it into <i>E. coli</i>, and tested the fluorescent output at different temperatures. These graphs show the relative fluorescent intensity of each thermometer between either 25 °C and 30 °C or 25 °C and 37 °C</p>
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<img src="https://2019.igem.org/wiki/images/a/a2/T--Rice--RNATcassette.svg" width = 60%>
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<p><b>Figure 1.</b>RNA thermometer testing circuit</p>
 
<img src="https://2019.igem.org/wiki/images/f/fe/T--Rice--BBa115003.png" width = 60%>
 
<img src="https://2019.igem.org/wiki/images/f/fe/T--Rice--BBa115003.png" width = 60%>
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<p><b>Figure 1.</b> Fluorescence intensity at 25 °C, 30 °C, and 37 °C.</p>
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<p><b>Figure 2.</b> Fluorescence intensity at 25 °C, 30 °C, and 37 °C.</p>
 
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Latest revision as of 00:28, 22 October 2019

RNA thermometer (PrfA)

An [http://2008.igem.org/Team:TUDelft/Temperature_analysis#RNA_thermometer RNA thermometer] that can be used for temperature sensitive post-transcriptional regulation which is based on a RNA thermometer from Listeria Monocytogenes that initiates translation at 37°C.

The image shows the secondary structure of the part after ligation to a protein coding part, as predicted by [http://rna.tbi.univie.ac.at/cgi-bin/RNAfold.cgi RNAfold]. Notice that the 3' end, including the scar and the start codon, does not belong to the part. The blue region is the Shine Dalgarno sequence which is the ribosome binding site. The blue nucleotides are the ones that had to be changed in order to get the desired secondary structure after introduction of the scar.

BBa K115003 ss.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization

To characterize this RNA thermometer in vivo, we made an RNA thermometer testing circuit, transformed it into E. coli, and tested the fluorescent output at different temperatures. These graphs show the relative fluorescent intensity of each thermometer between either 25 °C and 30 °C or 25 °C and 37 °C

Figure 1.RNA thermometer testing circuit

Figure 2. Fluorescence intensity at 25 °C, 30 °C, and 37 °C.