Team:Newcastle/End of crack & signalling system

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Revision as of 06:02, 14 June 2010

Subtilin

• A type 1 antimicrobial peptide [AMP] or lantibiotic produced by B. subtilis
  • (Lantibiotic = peptide-derived antibiotics with high antimicrobial activity against various Gram-positive bacteria, including pathogenic bacteria such as propionibacteria, staphylococci, clostridia, enterococci and streptococci.
  • NB. Subtilin  Subtilisin!!!!!
• Production of subtilin involves the spa gene cluster encompassing 10 genes, spaBTCSIFEGRK

The construct which contains:

• spaRK promotor
• spaR (subtilin peptide antibiotic Regulation) - the 220 amino acid product of this gene usually regulates the downstream production of subtilin antibiotic. It has an N-terminal domain that can be phosphorylated and a C-terminal domian that has DNA binding properties [1]
• spaK (subtilin peptide antibiotic Kinase) - this gene codes for a 325 amino acid histadine kinase peptide that phosphorylates the N-terminus of spaR [2]. This activates the DNA binding ability of the C-terminus of spaR, which in turn initiates transcription of the downstream gene. In the case of our construct, this gene is gfp.
• rrnO transcriptional terminator
• spaS promotor - a strong promotor inducible by upstream activation of spaRK. It can be placed in front any gene to regulate its activity.