Coding

Part:BBa_K801080:Design

Designed by: Alois Bruer   Group: iGEM12_TU_Munich   (2012-09-21)

Prepro-Thaumatin, yeast codon optimized


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]


Synthesized by IDT. The part is codon optimized for yeast. 3 forbidden restriction sites have been removed.

Please note, that the pre- and pro-part are split off, thus there is no possibility to add streptags or form other recombinant proteins.


Based on: http://www.ncbi.nlm.nih.gov/nuccore/121945717



Keywords: preprothaumatin, yeast, sweetner


Abbreviations:

Design Notes

Related BioBrick: -

Cloning details:

  • Designed in RFC10

Quality control measures:

  • Part was totally sequenced

Backbone:

  • Backbone name: pSB1C3
  • Resistance: Cp
  • Copynumber: high

Protein coding:

  • Protein: Preprothaumatin [Nucleotide 1 to 708]
  • The protein encoded is posttranslationally modified: Aa 1-22 and 229-236 are split off

Enzymatic activity: -

Cytotoxicity:
This Biobrick leads to the production of thaumatin. We investigated the toxicity of thaumatin and incubated yeast cells (strain INVSc) with 45 mM of thaumatin. We found, that thaumatin does not show any negative effect on the growth rate of yeast cells.

Safety notes:

Intellectual property: Information on patent situation: Preprothaumatin derived from Thaumatococcus daniellii

Corresponding part author/authors:

Source

Source:

  • Synthesized by idt


Organism:

  • Genesequence derived from Thaumatococcus daniellii
  • Codonoptimized for S. cerevisiae
  • Designed for the following Chassis: S. cerevisiae
  • Functionality in other chassis: Should work in other chassis as well (although optimized for S. cerevisiae)

References

Literature references:

  • [http://www.ncbi.nlm.nih.gov/pubmed/6327079 Pubmed: Edens L, Bom I, Ledeboer AM, Maat J, Toonen MY, Visser C, Verrips CT., 1984: Synthesis and processing of the plant protein thaumatin in yeast]
  • [http://www.ncbi.nlm.nih.gov/pubmed/19489607 Pubmed: Ide N, Sato E, Ohta K, Masuda T, Kitabatake N., 2009: Interactions of the sweet-tasting proteins thaumatin and lysozyme with the human sweet-taste receptor]
  • [http://www.ncbi.nlm.nih.gov/pubmed/17897626 Pubmed: Ide N, Masuda T, Kitabatake N., 2007: Effects of pre- and pro-sequence of thaumatin on the secretion by Pichia pastoris]

Database references:

  • [http://www.ncbi.nlm.nih.gov/nuccore/121945717 GenBank AB265690.1: Thaumatococcus daniellii mRNA for preprothaumatin I]
  • [http://http://www.ebi.ac.uk/interpro/IEntry?ac=IPR017949&query=thaumatin Interpro: Thaumatin, conserved site (IPR017949)]
  • [http://www.uniprot.org/uniprot/A1IIJ1 Uniprot: Preprothaumatin I]