Difference between revisions of "Part:BBa K2680260"

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<partinfo>BBa_K2680260 short</partinfo>
 
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3G eBFP2
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3G eBFP2 coding sequence flanked by prefix sticky end C and suffix sticky end D in a 3G-compatible backbone. For more information on 3G hybrid DNA assembly and 3G parts, see the [http://2018.igem.org/Team:William_and_Mary/Part_Collection William and Mary 3G Parts Page].
  
 
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===Usage and Biology===
 
===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
 
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<partinfo>BBa_K2680260 SequenceAndFeatures</partinfo>
 
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<partinfo>BBa_K2680260 parameters</partinfo>
 
<partinfo>BBa_K2680260 parameters</partinfo>
 
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===Description===
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This is an enhanced blue fluorescent protein with an excitation maximum of 363nm and an emission maximum of 448nm.<sup>[1]</sup> Engineered in Ai et al. 2007 through "rounds of random mutagenesis and screening."<sup>[2]</sup> Ai et al. 2007 describes eBFP2 as "a variant that is 4-fold brighter and 550-fold more photostable than EBFP."<sup>[2]</sup>.
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The paper does warn that eBFP2 "may retain wild-type GFP’s tendency to dimerize at high concentrations. However, it has been suggested that the A206V “superfolder” mutation of EBFP2 could hinder dimerization."<sup>[2]</sup>
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Two figures from Ai et al. 2007:
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<html><img src="https://static.igem.org/mediawiki/parts/5/56/T--William_and_Mary--eBFP2table.png" width="750px"/></html>
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<html><img src="https://static.igem.org/mediawiki/parts/8/8e/T--William_and_Mary--eBFP2graph.png" width="400px"/></html>
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Above: "Spectral characterization of new variants. Shown in each panel are the absorbance (green), excitation (black), and emission (red) spectra for the indicated protein."<sup>[2]</sup>
  
 
===References===
 
===References===
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[1] Subach OM, Cranfill PJ, Davidson MW, Verkhusha VV (2011) An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore. PLoS ONE 6(12): e28674. https://doi.org/10.1371/journal.pone.0028674
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[2] Ai, H., Shaner, N. C., Cheng, Z., Tsien, R. Y., & Campbell, R. E. (2007). Exploration of New Chromophore Structures Leads to the Identification of Improved Blue Fluorescent Proteins. Biochemistry, 46, 5904-5910. doi:10.1021/bi700199g
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2680260 SequenceAndFeatures</partinfo>

Latest revision as of 20:01, 16 October 2018


3G eBFP2

3G eBFP2 coding sequence flanked by prefix sticky end C and suffix sticky end D in a 3G-compatible backbone. For more information on 3G hybrid DNA assembly and 3G parts, see the [http://2018.igem.org/Team:William_and_Mary/Part_Collection William and Mary 3G Parts Page].

Description

This is an enhanced blue fluorescent protein with an excitation maximum of 363nm and an emission maximum of 448nm.[1] Engineered in Ai et al. 2007 through "rounds of random mutagenesis and screening."[2] Ai et al. 2007 describes eBFP2 as "a variant that is 4-fold brighter and 550-fold more photostable than EBFP."[2].

The paper does warn that eBFP2 "may retain wild-type GFP’s tendency to dimerize at high concentrations. However, it has been suggested that the A206V “superfolder” mutation of EBFP2 could hinder dimerization."[2]

Two figures from Ai et al. 2007:

Above: "Spectral characterization of new variants. Shown in each panel are the absorbance (green), excitation (black), and emission (red) spectra for the indicated protein."[2]

References

[1] Subach OM, Cranfill PJ, Davidson MW, Verkhusha VV (2011) An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore. PLoS ONE 6(12): e28674. https://doi.org/10.1371/journal.pone.0028674

[2] Ai, H., Shaner, N. C., Cheng, Z., Tsien, R. Y., & Campbell, R. E. (2007). Exploration of New Chromophore Structures Leads to the Identification of Improved Blue Fluorescent Proteins. Biochemistry, 46, 5904-5910. doi:10.1021/bi700199g

Sequence and Features


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 47
    Illegal BsaI.rc site found at 783