Difference between revisions of "Part:BBa K2970008"

Latest revision as of 02:11, 22 October 2019

Gate/Trigger test composition

To measure the functionality of our gate (BBa_K2970002, Figure 1B) and trigger (BBa_K2970000 and BBa_K2970001) we designed a test plasmid which contains both trigger and the gate, which is locking GFP, each embedded into a promoter and a terminator. The promoters of the trigger can be induced by either IPTG or arabinose to test different combinations of trigger concentrations. In contrast, the gate is translated constitutively.

The function of the gate is the regulation of the translation of a gene of interest, in this case GFP for testing, by formation of a hairpin by the mRNA that hides the ribosome binding site and the start codon. The mRNAs of the two triggers can form a complex due to complementarity (Figure 1A), which can open the gates hairpin and releases the ribosome binding site, thus translation can take place (Figure 2). The affinity of the trigger complex is higher than the affinity of the gate within. The gate can only be opened if both triggers are present, otherwise the complementary region is too short. If the gate is opened, GFP is expressed whose signal can be measured.

Results

Due to a high amount of secondary structures it was difficult to assemble this part. Therefore we could not provide characterization data.

Sequence and Features