Difference between revisions of "Part:BBa K4830027"

Revision as of 07:27, 12 October 2023

mCherry_C-terminus stop codon_Clover

The mCherry_C-terminus stop codon_Clover is protein coding sequence containing the mCherry fluorescent protein with a premature stop codon installed in the C-terminus before the Clover fluorescent protein. The presence of the premature stop codon prevents the expression of coding regions found downstream. This is particularly useful in assays such as the Traffic Light Reporter (TLR) assay.

Usage and Biology

The mCherry_C-terminus stop codon is utilized in the Traffic Light Reporter (TLR) assay, where a Clover fluorescent protein (commonly known as GFP) is found downstream. In this TLR assay, the Clover is not expressed due to the presence of the stop codon before the GFP. As such, the TLR assay is used to screen for DNA editing activity that swaps the stop codon into a functional amino acid, restoring the expression of Clover.

The entire coding sequence can be cloned into suitable plasmids and subsequently transfected into mammalian cells for in vitro DNA editing activity assay.

Sequence and Features

The sequence shows the mCherry fluorescent protein with a stop codon on the C-terminus. This is followed with a Clover fluorescent protein downstream.