Device

Part:BBa_K2463000

Designed by: Jack Lawrence, Balint Vidos   Group: iGEM17_Warwick   (2017-10-22)
Revision as of 19:58, 22 October 2017 by Psychosahedron (Talk | contribs) (Reformat)


Biofilm and Motility Regulation Device
The Biofilm and Motility Regulation Device is a single part capable of regulating motility of bacteria and dictyostelium. By overriding standard cellular processes via overexpression of enzymes, the system alters the biochemistry of the cell, specifically the expression of the signalling molecule responsible for regulating motility.

Background

  • Cyclic di-GMP is a secondary signalling molecule used to control motility in bacteria and dictyostelium: concentration of c-di-GMP alters motility.
  • c-di-GMP can be formed from two GTP molecules with the catalytic activity of a GGDEF domain found in diguanylate cyclases - in the case of the biofilm and motility control device, the diguanylate cyclase enzyme in question is PLeD.
  • c-di-GMP can be degraded into 5'-phosphoguanylyl(3'->5')guanosine (PGPG) with a water molecule and the catalytic activity of an EAL domain found in cyclic di-GMP phosphodiesterases - the c di-GMP phosphodiesterase in question is yhjH gene.

Function

The device contains a not-gate function between the PLeD and yhjH genes (Mnt repressor -> terminator -> repressible promoter). When the device is not being transcribed the yhjH gene is upregulated, causing c-di-GMP within the cell to be degraded. When the device is transcribed the yhjH gene is silenced and the PLeD protein is produced, causing the synthesis of c-di-gmp. The yhjH gene lacks a degrading tag thus the cells take a longer time to respond to the device being activated (exact details are unknown).

Usage

The device was designed to provide a simplified system of biofilm and motility manipulation. Insert a promoter downstream of the device: the rate of transcription of the device defines the function. For motility do not transcribe the device, for biofilm formation upregulate transcription of the device. Place the device in a circuit in order to override cellular responses regarding motility and biofilm formation.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1806
    Illegal BamHI site found at 32
    Illegal BamHI site found at 1190
    Illegal XhoI site found at 1768
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 448
    Illegal NgoMIV site found at 595
    Illegal AgeI site found at 931
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 105


[edit]
Categories
Parameters
None