Difference between revisions of "Part:BBa K1041000:Design"

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===Design Notes===
 
===Design Notes===
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An NdeI restriction site was added at the start of the RFP coding region of [[BBa_J04450]] using mutagenesis.
+
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 we used to create part [[Bba_K1041002]].
  
 
===Source===
 
===Source===
−
 
+
Bba_K1041000 is an improvement on the part Bba_J04450.
−
[[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===
 
===References===
 +
1. Campbell, R., Tour, O., Palmer, A., Steinbach, P., Baird, G., Zacharias, D & Tsien, R (2002) ''A Monomeric Red Fluorescent Protein''Proc 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 conditions''FEBS 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

Latest revision as of 21:41, 4 October 2013


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 we used to create 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