Difference between revisions of "Part:BBa K2970003"

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<partinfo>BBa_K2970003 short</partinfo>
 
<partinfo>BBa_K2970003 short</partinfo>
  
Together with Trigger 2 (<partinfo>BBa_K2970001</partinfo>), it forms a construct that can open the gate (<partinfo>BBa_K2970002</partinfo>).
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Trigger 1 forms together with Trigger 2 (<partinfo>BBa_K2970001</partinfo>) a complex due to complementary regions. This complex can open a gate (<partinfo>BBa_K2970002</partinfo>) to enable translation of a gene that is locked with this gate to regulate translation. After binding of the trigger complex due to complementarity, the hairpin of the gate will open and release the ribosome binding site and the start codon, thus translation can occur.  
The affinity between the trigger complex and the gate is greater than that of the gate to itself (loop). A single trigger cannot open the gate because it has only half the required complementary sequence.
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We used this system to regulate a chloramphenicol resistance, which is locked by the gate.
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To transform the trigger into bacteria it should be put on a plasmid together with a promoter and a terminator as we did with this part.
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After transformation of both trigger plasmids and the gate plasmid in one bacterium all three mRNA structures will be formed, the gate will be opened, and the translation of the chloramphenicol resistance can take place. Bacteria that took all three plasmids are able to survive on media with chloramphenicol. This system can be used to transform many genes of interest on three different plasmids into bacteria with only using one antibiotic resistance instead of three different resistances.
  
  

Revision as of 11:37, 20 October 2019


Trigger 1 Composition

Trigger 1 forms together with Trigger 2 (BBa_K2970001) a complex due to complementary regions. This complex can open a gate (BBa_K2970002) to enable translation of a gene that is locked with this gate to regulate translation. After binding of the trigger complex due to complementarity, the hairpin of the gate will open and release the ribosome binding site and the start codon, thus translation can occur.

We used this system to regulate a chloramphenicol resistance, which is locked by the gate. To transform the trigger into bacteria it should be put on a plasmid together with a promoter and a terminator as we did with this part. After transformation of both trigger plasmids and the gate plasmid in one bacterium all three mRNA structures will be formed, the gate will be opened, and the translation of the chloramphenicol resistance can take place. Bacteria that took all three plasmids are able to survive on media with chloramphenicol. This system can be used to transform many genes of interest on three different plasmids into bacteria with only using one antibiotic resistance instead of three different resistances.


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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]