Part:BBa_K218006:Experience

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=Applications of BBa_K218006

This signalling circuit encodes the proteins necessary for the V. harveyi signalling cascade. Once suitable expression levels of LuxPQ have been established based on the library of ∆σ70, this circuit will be fully functional. The power of the AI-2 signalling system coupled with a response by using the qrr4 promoter (Part:BBa_K131017) in a laboratory strain of E. coli lies within the principle of QS and coordinating bacterial behaviour. The E. coli will be able to express a gene with a desired function at high population densities simply if this gene is cloned downstream of the qrr4 promoter. This engineered system will prove quite versatile will respect to the output from AI-2 signalling.

The usefulness of AI-2 signalling in E. coli stretches far beyond the ability to clone in a gene of interest to express in high population densities. This safe, non-pathogenic, laboratory E. coli model of AI-2 signaling will contribute to the understanding of QS systems used by pathogenic bacteria to induce virulence. Periplasmic protein LuxP has been identified as a ribose-binding protein and its crystal structure has been determined along with important amino acids for AI-2 binding. Although this allows for the investigation of other LuxP-binding molecules in silico, the presence of AI-2 signalling in E. coli will allow for in vitro investigations of activating and inactivating ligands. With this comes the development of novel therapeutics aimed at attenuating QS and thus virulence induction in pathogenic bacteria by, for example, synthesizing a drug capable of competitively inhibiting the AI-2 binding site on LuxP. Targeting and preventing QS is particularly appealing considering the emergence of increasingly antibiotic resistant bacteria.

User Reviews

We are Team-IIT Madras 2015, we needed this biobrick for our project. We have found out that this biobrick contains some errors, which are at 1125-1127bp [TGA] is a stop codon for LuxP, while at 1127-1129bp [ATG] is the start codon for LuxQ protein. LuxQ protein will not be produced because of frameshift. Also, at 5422-5424bp [TGA] is the stop codon for LuxU protein and at 5421-5423 bp [ATG] is the start codon for LuxO protein. Because of frameshift , we would not get LuxU protein.

UNIQ351931485fedf3ea-partinfo-00000000-QINU UNIQ351931485fedf3ea-partinfo-00000001-QINU