Coding

Part:BBa_K4159007

Designed by: Melissa Hendrén   Group: iGEM22_Aalto-Helsinki   (2022-07-14)


AgrC (S. epidermidis)

Accessory gene regulator protein C (AgrC) in S. epidermidis bacteria. Is a part of the quorum sensing gene complex. Encodes for the receptor where AIP signaling molecules binds.

Description: This is the gene that encodes for the accessory gene-regulating protein C (AgrC). The protein works as a receptor, that binds the signaling peptide AIP-1. When AIP-1 binds to the receptor, 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 AgrC is a part of the quorum sensing system which can be found in the Stapholycoccus bacteria family, and this specific AgrC 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.

Fig.1 Schematic picture of the quorum sensing system in gram-positive bacteria (Rutherford, 2012)

Characterization: This part will be a part of the detecting mechanism which also includes the AgrA, 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 AgrC 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.

https://www.genome.jp/entry/ser:SERP1492


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None