Difference between revisions of "Part:BBa K4169011"

 
 
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Based on the characteristics of secondary structure changes of RNA under different temperature conditions, the RNA thermometer element was introduced in this project. An RC sequence is present on this RNA thermometer, which recruits RNase E to degrade subsequent RNA fragments. At low temperatures,such as 27℃, RNA thermometers will maintain the secondary stem-loop structure, and RC sequences will complement and pair with other bases, unable to recruit RNase, so subsequent sequences will be expressed normally. However, at 37℃, the secondary structure opens, and the subsequent RNA fragments are cleaved and degraded by RNA enzymes.In order to change the temperature at which it functions, we mutated its gene sequence, that is, the first A after ARC was changed to T.
 
Based on the characteristics of secondary structure changes of RNA under different temperature conditions, the RNA thermometer element was introduced in this project. An RC sequence is present on this RNA thermometer, which recruits RNase E to degrade subsequent RNA fragments. At low temperatures,such as 27℃, RNA thermometers will maintain the secondary stem-loop structure, and RC sequences will complement and pair with other bases, unable to recruit RNase, so subsequent sequences will be expressed normally. However, at 37℃, the secondary structure opens, and the subsequent RNA fragments are cleaved and degraded by RNA enzymes.In order to change the temperature at which it functions, we mutated its gene sequence, that is, the first A after ARC was changed to T.
  
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===Usage and Biology===
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===Design and Mutation===
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<p>We designed the mutation primers <b>(Figure 1)</b>and the expected mutation sequences<b>(Figure 2)</b></p>
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<center><img src="https://static.igem.wiki/teams/4169/wiki///rna-t/a-rna-thermometer-mutation-2.png
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<center><b>Figure 1. </b> the mutation primers</center>
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<center><img src="https://static.igem.wiki/teams/4169/wiki///rna-t/a-rna-thermometer-mutation-2-2.png
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<center><b>Figure 2. </b> the expected mutation sequences are highlighted in red</center>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4169011 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K4169010 SequenceAndFeatures</partinfo>
  
  
 
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K4169011 parameters</partinfo>
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<partinfo>BBa_K4169010 parameters</partinfo>
 
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==Reference==
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<p>Hoynes-O'Connor A, Hinman K, Kirchner L, Moon TS. De novo design of heat-repressible RNA thermosensors in E. coli. Nucleic Acids Res. 2015 Jul 13;43(12):6166-79. </p>

Latest revision as of 10:37, 13 October 2022


A RNA thermometer mutation 2

Based on the characteristics of secondary structure changes of RNA under different temperature conditions, the RNA thermometer element was introduced in this project. An RC sequence is present on this RNA thermometer, which recruits RNase E to degrade subsequent RNA fragments. At low temperatures,such as 27℃, RNA thermometers will maintain the secondary stem-loop structure, and RC sequences will complement and pair with other bases, unable to recruit RNase, so subsequent sequences will be expressed normally. However, at 37℃, the secondary structure opens, and the subsequent RNA fragments are cleaved and degraded by RNA enzymes.In order to change the temperature at which it functions, we mutated its gene sequence, that is, the first A after ARC was changed to T.

Design and Mutation

We designed the mutation primers (Figure 1)and the expected mutation sequences(Figure 2)

无标题文档

Figure 1. the mutation primers
Figure 2. the expected mutation sequences are highlighted in red

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 39


Reference

Hoynes-O'Connor A, Hinman K, Kirchner L, Moon TS. De novo design of heat-repressible RNA thermosensors in E. coli. Nucleic Acids Res. 2015 Jul 13;43(12):6166-79.