Team:Newcastle/End of crack & signalling system

From 2010.igem.org

(Difference between revisions)
(Production and Immunity Bricks)
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*''spaI'' 's gene product sequesters subtilin.
*''spaI'' 's gene product sequesters subtilin.
*''spaFEG'' genes code for transporters.
*''spaFEG'' genes code for transporters.
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[[Image:spa]]
=Genes required for swarming=
=Genes required for swarming=

Revision as of 15:37, 2 July 2010

Contents

Subtilin

2008Brick

  • A type 1 antimicrobial peptide [AMP] or lantibiotic produced by B. subtilis
    • NB. 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 not same as Subtilisin!!!!!
  • Production of subtilin involves the spa gene cluster encompassing 10 genes, spaBTCSIFEGRK

NCL - Biobrick for Subtilin Sensor.PNG

The construct above 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.

Production and Immunity Bricks

  • spaBTCS genes are required for production.
  • spaIFEG genes are required for immunity.
  • spaB and C are modifiers of the subtilin precursor.
  • spaT is a transporter.
  • spaS is the gene coding for the subtilin precursor peptide.
  • spaI 's gene product sequesters subtilin.
  • spaFEG genes code for transporters.

File:Spa

Genes required for swarming

  1. comP: Histidine sensor kinase of ComX and phrC
  2. comA: Response regulator of Comp
  3. sfp: Active surfactin synthetase
  4. srfA,B and C: Surfactin synthetase
  5. swrA,B and C: Novel gene for swarming and not for swimming
  6. efp: elongation factor P
  7. yabR and ymfl: Contribute to proper coordinationamong cells of the swarming population but are not absolutely required for surface migration.
  8. cheC and D: chemotaxis protein
  9. sigD: transcription factor sigma D, which is required for the expression of genes activated in late phase of flagellum biosynthesis.