Tag

Part:BBa_K208005:Design

Designed by: USU iGEM 2009   Group: iGEM09_Utah_State   (2009-10-12)

TorA Signal Peptide - Silver Fusion Compatible


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]


Design Notes

This part was designed for Silver-fusion compatibility. It is in plasmid pSB3K3.

Another TorA BioBrick was constructed by the 2008 Wisconsin iGEM team. This part, however, is not fusion-compatible. Accordingly, our team decided to design a Silver-fusion compatible part. Additionally, the sequence that we chose to use is different than that used by the Wisconsin team. Based on a number of references (Palmer, 2004; NCBI, Institute for Biomolecular Design), we selected a 126 bp sequence. Analysis of the existing Wisconsin-designed TorA sequence by a simple BLAST search on NCBI yields no significant matches, so we are not aware of the origin of their sequence. The 2009 IBB Pune iGEM team is deriving a fusion-compatible TorA sequence from the part originally designed by Wisconsin 2008 (see BBa_K233307). For this purpose, they selected to not use the Silver-fusion method. Accordingly, the addition of our Silver-fusion TorA BioBrick part adds an alternative for studying protein secretion.

Source

This part was designed to have the Silver-fusion prefix and suffix, and was synthetically produced by DNA 2.0. The obtained plasmid was then cut with EcoRI and SpeI. The insert was then ligated with a BioBrick compatible vector, pSB3K3. This part has been sequenced.

References

1. Barrett CML, Ray N, Thomas JD, Robinson C, Bolhuis A (2003) Quantitative export of a reporter protein, GFP, by the twin-arginine translocation pathway in Escherichia coli. 2. Biochem BIophys Res Comm 304:279-284
3. Berks BC (1996) Mol Microbiol 22:393-404
4. Berks BC, Sargent F, Palmer T (2000) The Tat protein export pathway. Mol Microbiol 35:260-274
5. Buchanan G (2008) Features of a twin-arginine signal peptide required for recognition by a Tat proofreading chaperone. FEBS letters 582:3979
6. Choi JH, Lee SY (2004) Secretory and extracellular production of recombinant proteins using Escherichia coli Appl Microbiol Biotechnol 64:625-635
7. http://www.ncbi.nlm.nih.gov/protein/806336?from=1&to=39&report=gpwithparts
8. Palmer T, Berks BC (2004) The Tat protein export pathway In: Oudega B, editor. Kluwer Academic Publishers, pp 51-64
9. Santini C, Bernadac A, Zhang M, Chanal A, Ize B, Blan co C, Wu L (2001) Translocation of jellyfish green fluorescent protein via the Tat system of Escherichia coli and change of its periplasmic localization in response to osmotic up-shock. J Biol Chem 276:8159-8164
10. Thomas JD, Daniel RA, errington J, Robinson C (2001) Export of Active green fluorescent protein to the periplasm by the twin-arginine translocase (Tat) pathway in Escherichia coli. Mol Microbiol 39:47-53