Difference between revisions of "Part:BBa K3084000"
 
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<partinfo>BBa_K3084000 short</partinfo>
 
<partinfo>BBa_K3084000 short</partinfo>
  
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This Venus YFP gene has been designed for optimal translational efficiency from the genome of bacteriophage S-TIP37 during infection of its host, Synechococcus sp. strain WH8109. The translation of this gene is the same as the Venus YFP gene (NCBI accession number: ACQ43942). To maximise translational efficiency during S-TIP37 infection, the codon distribution of the S-TIP37 capsid gene, putatively the most strongly expressed gene on the S-TIP37 genome, was applied to the Venus YFP gene.
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This Venus YFP gene has been designed for optimal translational efficiency during expression from the genome of bacteriophage S-TIP37 when infecting its host, Synechococcus sp. strain WH8109. The translation of this gene is the same as the Venus YFP gene (NCBI accession number: ACQ43942). To maximise translational efficiency during S-TIP37 infection, the codon distribution of the S-TIP37 capsid gene, putatively the most strongly expressed gene on the S-TIP37 genome, was applied to the Venus YFP gene.
  
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Comparing the translational efficiency to other YFPs in the S-TIP37 expression system required an experimental setup that was too complicated for the scope of our summer iGEM project. However, the fluorescence of our codon optimised gene was successfully tested using the pET expression system in E. Coli. Proof of fluorescence is demonstrated in figure 1.
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Comparing the translational efficiency to other YFPs in the S-TIP37 expression system required an experimental setup that was too complicated for the scope of our iGEM project. However, the fluorescence of our codon optimised gene was successfully tested using the pET expression system in E. Coli. Proof of fluorescence is demonstrated in figure 1.
  
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[[File:T--KU LEUVEN--Fluorescence_v2.jpg|440px]]
  
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[[File:T--KU LEUVEN--Fluorescence Venus.jpg|440px]]
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Figure 1. Colonies showing the fluorescence of the <partinfo>K3084000</partinfo> Venus YFP part on UV table.
  
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Optimal excitation wavelengths were determined by Sarkar et al. and found to be: 285 and 470 nm.  
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Optimal excitation wavelengths were determined by Sarkar et al. (2009) and found to be: 285 and 470 nm.  
  
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===Usage and biology===
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<span class='h3bb'>Usage and biology</span>
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This part is useful as a reporter, most suitable for expression during Synechococcus infection by phage S-TIP37.
 
This part is useful as a reporter, most suitable for expression during Synechococcus infection by phage S-TIP37.
  
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===Sequence and Features===
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<span class='h3bb'>References</span>
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[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2754229/] Sarkar, P., Koushik, S. V., Vogel, S. S., Gryczynski, I., & Gryczynski, Z. (2009). Photophysical properties of Cerulean and Venus fluorescent proteins. Journal of biomedical optics, 14(3), 034047. doi:10.1117/1.3156842
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3084000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3084000 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K3084000 parameters</partinfo>
 
<partinfo>BBa_K3084000 parameters</partinfo>
 
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===References===
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[https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2754229/] Sarkar, P., Koushik, S. V., Vogel, S. S., Gryczynski, I., & Gryczynski, Z. (2009). Photophysical properties of Cerulean and Venus fluorescent proteins. Journal of biomedical optics, 14(3), 034047. doi:10.1117/1.3156842

Latest revision as of 16:51, 21 October 2019

Venus Yellow Fluorescent Protein (YFP), S-TIP37 codon optimised

This Venus YFP gene has been designed for optimal translational efficiency during expression from the genome of bacteriophage S-TIP37 when infecting its host, Synechococcus sp. strain WH8109. The translation of this gene is the same as the Venus YFP gene (NCBI accession number: ACQ43942). To maximise translational efficiency during S-TIP37 infection, the codon distribution of the S-TIP37 capsid gene, putatively the most strongly expressed gene on the S-TIP37 genome, was applied to the Venus YFP gene.

Comparing the translational efficiency to other YFPs in the S-TIP37 expression system required an experimental setup that was too complicated for the scope of our iGEM project. However, the fluorescence of our codon optimised gene was successfully tested using the pET expression system in E. Coli. Proof of fluorescence is demonstrated in figure 1.

T--KU LEUVEN--Fluorescence v2.jpg

Figure 1. Colonies showing the fluorescence of the BBa_K3084000 Venus YFP part on UV table.

Optimal excitation wavelengths were determined by Sarkar et al. (2009) and found to be: 285 and 470 nm.

Usage and biology

This part is useful as a reporter, most suitable for expression during Synechococcus infection by phage S-TIP37.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 424
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 682
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[1] Sarkar, P., Koushik, S. V., Vogel, S. S., Gryczynski, I., & Gryczynski, Z. (2009). Photophysical properties of Cerulean and Venus fluorescent proteins. Journal of biomedical optics, 14(3), 034047. doi:10.1117/1.3156842