RNA

Part:BBa_K2970002

Designed by: Marie-Theres Thieme   Group: iGEM19_Hamburg   (2019-10-02)
Revision as of 01:35, 22 October 2019 by MarieTT (Talk | contribs)


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 structure that hides the ribosome binding site and start codon of the gene of interest, thus translation can not be initiated (Figure 1B). A complementary part to the gate, a so called trigger, is needed to open the hairpin and release the ribosome binding site. In our 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 hairpin). 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).

===Results===

For results visit page of BBa_K2980006. 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]


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