Difference between revisions of "Part:BBa K2978100"

Revision as of 05:18, 21 October 2019

Auto Inducer Protein for QS in Clostridioides difficile

This composite part was constructed to reproduce the Quorum Sensing signaling of C. difficile. It´s mechanisms is based on the Lubkowicz, et al., (2018) assays who reproduces the QS signaling from S. aureus.
Naturally, in C. difficile AgrD protein is code in higher expression levels when conditions are favorable for the pathogen. This peptide is modified and transport to the medium by the transmembrane AgrB protein, such modification results in the auto inducer protein (AIP) that recreates an infection/colonization induction in the bacteria.
AIP is sensing by the AgrC membrane protein, which phosphorylate AgrA protein when AIP is detected in the outsider of the bacteria. Once two AgrA protein are phosphorylated creates a dimer protein, and this joining form a DNA binding domain, still unknows in C. difficile, but well reported in S. aureus.
Both enzymes in this part are stablished with the objective to produce the AIP when [http://openwetware.org/wiki/IPTG IPTG] is applied. Also, a GFP reporter is code polycistronically after the AgrD peptide, as a way to indirectly quantify the AIP, because methods like nickel affinity chromatography are not useful in this cases for the short size of the peptide, and the possible interference that a HisTag can make when AgrD is processed by AgrB.

Sequence and Features