Difference between revisions of "Part:BBa K2970002"
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This gate is a toehold switch system with which a gene of interest can be locked and regulated on a translational level using mRNA as regulator. After transcription, the mRNA of this gate forms a hairpin that hides the ribosome binding site and start codon of the gene of interest, thus translation can not occur. A complementary part to the gate (trigger) is needed to open the hairpin and release the ribosome binding site. In this case two triggers are needed that form a trigger complex to open the gate (<partinfo>BBa_K2970000</partinfo> and <partinfo>BBa_K2970001</partinfo>). The affinity between the trigger complex and the gate is greater than that of the gate to itself (in the loop). A single trigger cannot open the gate because it has only half the required complementary sequence.
 
This gate is a toehold switch system with which a gene of interest can be locked and regulated on a translational level using mRNA as regulator. After transcription, the mRNA of this gate forms a hairpin that hides the ribosome binding site and start codon of the gene of interest, thus translation can not occur. A complementary part to the gate (trigger) is needed to open the hairpin and release the ribosome binding site. In this case two triggers are needed that form a trigger complex to open the gate (<partinfo>BBa_K2970000</partinfo> and <partinfo>BBa_K2970001</partinfo>). The affinity between the trigger complex and the gate is greater than that of the gate to itself (in the loop). A single trigger cannot open the gate because it has only half the required complementary sequence.
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<img width="100%" src="https://2019.igem.org/wiki/images/b/bc/T--Hamburg--PartsFigure11.jpeg">
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<figcaption>
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<b>Figure 1: </b>A) Formation of trigger complex after translation. B) mRNA of gate sequence forms secondary structures that hide the ribosome binding site and the start codon.
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<html>
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<figure>
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<img width="90%" src="https://2019.igem.org/wiki/images/4/49/T--Hamburg--Part_Figure6.jpg">
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<b>Figure 2: </b>Opening of the gate due to annealing of trigger complex to gate.
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</figure>
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To transform this part into bacteria it should be put on a plasmid together with a promoter and a terminator. We used this part to regulate a chloramphenicol resistance gene that we put behind the gate (<partinfo>BBa_K2970006</partinfo>).
 
To transform this part into bacteria it should be put on a plasmid together with a promoter and a terminator. We used this part to regulate a chloramphenicol resistance gene that we put behind the gate (<partinfo>BBa_K2970006</partinfo>).
  

Revision as of 09:11, 21 October 2019


Gate

This gate is a toehold switch system with which a gene of interest can be locked and regulated on a translational level using mRNA as regulator. After transcription, the mRNA of this gate forms a hairpin that hides the ribosome binding site and start codon of the gene of interest, thus translation can not occur. A complementary part to the gate (trigger) is needed to open the hairpin and release the ribosome binding site. In this case two triggers are needed that form a trigger complex to open the gate (BBa_K2970000 and BBa_K2970001). The affinity between the trigger complex and the gate is greater than that of the gate to itself (in the loop). A single trigger cannot open the gate because it has only half the required complementary sequence.

Figure 1: A) Formation of trigger complex after translation. B) mRNA of gate sequence forms secondary structures that hide the ribosome binding site and the start codon.
Figure 2: Opening of the gate due to annealing of trigger complex to gate.
To transform this part into bacteria it should be put on a plasmid together with a promoter and a terminator. We used this part to regulate a chloramphenicol resistance gene that we put behind the gate (BBa_K2970006).


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]