Part:BBa_K1041000:Design

RFP Coding Device

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 781
  Illegal AgeI site found at 893
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Team NRP-UEA_Norwich 2013 improved BBa_J04450 by adding an NdeI restriction site at the start of the RFP coding region of using mutagenesis. This was to allow either the promoter region or RFP gene to be excised and exchanged for a different promoter or gene and provide restriction sites for further cloning such as part Bba_K1041002.

Source

Bba_K1041000 is an improvement on the part Bba_J04450. This part contains the gene for the highly engineered version of the red fluorescent protein from Discosoma coral (DsRed). Most fluorescent proteins form a quarternary structure including Green florescent protein (GFP) and the naturally occuring DsRED [1]. Unlike GFP, the final structure of DsRED has hindered its ability to be a useful tool in biotechnology [2] as it forms intracellular aggregates [3]. The evolution of DsRED to a monomeric Red Fluorescent Protein provides an alternative to GFP as a reporter [1].

References

1. Campbell, R., Tour, O., Palmer, A., Steinbach, P., Baird, G., Zacharias, D & Tsien, R (2002) A Monomeric Red Fluorescent ProteinProc Natl Acad Sci U S A, Volume:99,Issue:12, pp:7877-7882 2.Vrzheshch, V., Akovbian, A., Varfolomeyev, D & Verkhusha, V (2000) Denaturation and partial renaturation of a tightly tetramerized DsRed protein under mildly acidic conditionsFEBS Lett, Volume:487, Issue:2, pp:203-208 3.Lauf, U., Lopez, P & Falk, M (2001)Expression of fluorescently tagged connexins: a novel approach to rescue function of oligomeric DsRed-tagged proteins FEBS Lett, Volume:498, Issue:1, pp:11–15