Difference between revisions of "Part:BBa K3725040"

Revision as of 01:30, 21 October 2021

T7 Fusarium Trigger

Overview

The T7 Phytophthora Trigger is designed to be used in conjunction with the T7 promoter to induce GFP expression for Lambert iGEM’s P. Cryptogea toehold biosensor. When the trigger RNA sequence is present, it binds to the complementary sequence in the toehold switch and unravels the hairpin loop allowing the reporter protein (GFP) to be expressed, producing green fluorescence. The sequence was designed via NUPACK using an input code provided by Takahashi et. al. We ordered the insert in a pUCIDT Kan plasmid from Integrated DNA Technologies.

Description

Toehold biosensors, which are composed of a switch and trigger, are highly orthogonal riboregulators that activate translation in response to a specific RNA sequence. The switch is composed of a hairpin loop structure that represses translation through its complementary bases in between the ribosomal binding site and the start codon, which is followed by a linker sequence. Once the toehold is exposed to the trigger sequence, the complementary base pairs on the trigger will bind to the toehold, which exposes the ribosomal binding site. RNA polymerase can then bind to the RBS and initiate translation of the reporter protein.