Difference between revisions of "Part:BBa K3487007"
 
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===Description===
 
===Description===
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Usually the suicide switch that prevents organisms from escaping will be set to auxotrophic type, which requires gene resection technology, and whether it is accurate is a big problem that needs to be faced. The riboswitch does not have these problems. The gene sequence is directly introduced into the required vector, and the switch directly forms a multiple stem-loop structure on the mRNA to stop translation.
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Riboswitch is a regulatory part of an mRNA molecule that can specifically bind a metabolite and regulate gene expression. Till now, more than 20 riboswitches have been discovered. Among them, only two amino acids riboswitches were found: a lysine riboswitch and a glycine riboswitch. Several previous studies examined the functionality and manipulation of the B. subtilis lysine riboswitch. A lysine riboswitch was also found in <i>E. coli</i>, which revealed in fact the mechanism of lysine feedback repression of aspartkinase III in <i>E. coli</i>.
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[[File:T--SZPT-CHINA--RS.png|900px|center|]]
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The following is the control principle diagram
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[[File:T--SZPT-CHINA--RS.png|500px|center|]]
 
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===Usage and Biology===
 
===Usage and Biology===
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===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K3487007 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3487007 SequenceAndFeatures</partinfo>
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==2020 SZPT-CHINA==
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===Result===
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In our project, we set up a suicide mechanism regulated by this lysine riboswitch part. When the bacteria colonize the oral cavity, the riboswitch controlled by lysine is turned off. When the bacteria escape the oral environment, the riboswitch opens and expresses the toxic protein RalR, which is a non-specific endonuclease.
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In order to determine at what concentration of lysine this switch can be turned on. We connected sfGFP to lysine riboswitch and then tested the fluorescent expression in different concentrations of lysine for 24 hours. The result is as follows.
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[[File:T--SZPT-CHINA--RS2.png|700px|center]]
  
 
===References===
 
===References===
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【2】Garst AD, Héroux A, Rambo RP, Batey RT. Crystal structure of the lysine riboswitch regulatory mRNA element. J Biol Chem. 2008;283(33):22347-22351.
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Garst AD, Héroux A, Rambo RP, Batey RT. Crystal structure of the lysine riboswitch regulatory mRNA element. J Biol Chem. 2008;283(33):22347-22351.
 
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K3487007 parameters</partinfo>
 
<partinfo>BBa_K3487007 parameters</partinfo>
 
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Latest revision as of 14:56, 27 October 2020


Lysine riboswitch

Description

Riboswitch is a regulatory part of an mRNA molecule that can specifically bind a metabolite and regulate gene expression. Till now, more than 20 riboswitches have been discovered. Among them, only two amino acids riboswitches were found: a lysine riboswitch and a glycine riboswitch. Several previous studies examined the functionality and manipulation of the B. subtilis lysine riboswitch. A lysine riboswitch was also found in E. coli, which revealed in fact the mechanism of lysine feedback repression of aspartkinase III in E. coli. The following is the control principle diagram

T--SZPT-CHINA--RS.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 12
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 284
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 256

2020 SZPT-CHINA

Result

In our project, we set up a suicide mechanism regulated by this lysine riboswitch part. When the bacteria colonize the oral cavity, the riboswitch controlled by lysine is turned off. When the bacteria escape the oral environment, the riboswitch opens and expresses the toxic protein RalR, which is a non-specific endonuclease. In order to determine at what concentration of lysine this switch can be turned on. We connected sfGFP to lysine riboswitch and then tested the fluorescent expression in different concentrations of lysine for 24 hours. The result is as follows.

T--SZPT-CHINA--RS2.png

References

Garst AD, Héroux A, Rambo RP, Batey RT. Crystal structure of the lysine riboswitch regulatory mRNA element. J Biol Chem. 2008;283(33):22347-22351.