Team:Heidelberg/Favorite Parts

From 2013.igem.org

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Revision as of 21:07, 28 October 2013

Favorite Parts. Now it is Your turn. Start working with NRPS!


Best BioBrick Part or Device, Natural.

BBa_K1152013. IndC Blue Pigment Production Device.

  • The indigoidine synthetase IndC is a single-module NRPS
  • Catalyzes the formation of the blue pigment indigoidine by circularizing glutamine
  • Indigoidine is visible to the naked eye
  • Indigoidine production can easily be quantified photometrically and is thus an ideal reporter
  • In constrast to many other pigment production devices, IndC does not need any external substrate supply

    • Best BioBrick Part or Device, Engineered.

    BBa_K1152007. The GFP for Non-Ribosomal Peptides.

  • Enables labeling of custom non-ribosomal peptides with a blue pigment in vivo (see Indigoidine-Tag page)
  • NRPS modules of choice are introduced in front of an engineered IndC module containing an additional C-domain for communication with the previous module
  • ccdB is replaced during cloning; close to 100 % cloning efficiency
  • Peptide production can be seen with the naked eye or quantified photometrically
  • Labelled peptides can easily be purified and characterized by thin-layer chromatography
  • Helper-construct for high-throughput cloning of custom NRPSs for production of synthetic peptides (see RFC 100 for details)

    • Most improved Registry Part.

    BBa_K1152009. Highly-Efficient NRPS-activating PPTase.

  • Helper plasmid for efficient peptide production by engineered or natural NRPSs
  • Transfers a 4'-PPT residue from CoA to a conserved serine residue in the T-Domain of NRPS modules, thereby activating the NRPS module
  • Improves parts BBa_K802006 and BBa_K302010 which are PPTases that were present in the registry, but neither annotated nor characterized as such

    • Best Part Collection.

    BBa_K1152015 - 2019. A Collection of Synthetic Indigoidine Synthetases Engineered by T-domain Exchange of IndC.

  • T-domain is important for NRPS efficiency
  • IndC was used as scaffold and the native IndC T-domain was exchanged by different natural and synthetic T-domains
  • Differing in indigoidine production efficiency
  • BBa_K1152015 outperforms the native IndC (BBa_K1152013) when combined with the Sfp or EntD PPTase (red square)
  • Collection can now be applied for tagging synthetic peptides produced by engineered NRPSs (see Indigoidine-Tag subproject)
    • Thanks to

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