Difference between revisions of "Part:BBa K3084000"
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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. | 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. | ||
| − | [[File:T--KU LEUVEN-- | + | [[File:T--KU LEUVEN--Fluorescence_v2.jpg|440px]] |
Figure 1. Colonies showing the fluorescence of the <partinfo>K3084000</partinfo> Venus YFP part under excitation light. | Figure 1. Colonies showing the fluorescence of the <partinfo>K3084000</partinfo> Venus YFP part under excitation light. | ||
Revision as of 15:05, 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.
Figure 1. Colonies showing the fluorescence of the BBa_K3084000 Venus YFP part under excitation light.
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 424
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 682
- 1000COMPATIBLE 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