Part:BBa_K4159008:Design

Description: This is the gene that encodes for the accessory gene-regulating protein A (AgrA). The protein works as the internal signaling protein AgrA. When AIP-1 signaling peptide binds to the receptor (AgrC), the internal signaling protein AgrA will be phosphorylated at the receptor site, and then activates the P2 promoter which starts the downstream gene transcription (Rutherford, 2012). The AgrA is a part of the quorum sensing system which can be found in the Stapholycoccus bacteria family, and this specific AgrA gene is derived from the S. epidermidis bacteria. The choice of this bacteria was because it is one of the biofilm producing bacteria in chronic wounds, and additionally it has strains of biosafety level 1. The exact gene sequence was derived from the Kegg database.

Characterization: This part will be a part of the detecting mechanism which also includes the AgrC, and GFP encoding genes. The AgrC and AgrA will be cloned into the plasmid, providing the E.coli with the needed proteins to detect the AIP-1’s in the environment and activate P2 which has the GFP downstream. The part was ordered as a gene fragment in combination with also other parts, and as the ordered fragment was then more complex, some codon optimization for the AgrA was done through the gene fragment producing company, IDT.

References: [1] Rutherford ST, Bassler BL. Bacterial quorum sensing: its role in virulence and possibilities for its control. Cold Spring Harb Perspect Med. 2012 Nov 1;2(11):a012427. doi: 10.1101/cshperspect.a012427. PMID: 23125205; PMCID: PMC3543102. [2] https://www.genome.jp/entry/ser:SERP1493