Part:BBa_K4813000
dTomato fluoresence protein Codon Optimized for E coli
This part encodes a protein called dTomato, which is a red fluorescence protein. It forms a dimer and has a molecular weight of around 27.0 kDa. This protein originates from a species called Discosoma [1].
To enhance its performance for our project, we used IDT codon optimization tools to optimize the sequence. We incorporated this optimized sequence into two composite parts. One part served as a reporter gene for detecting formaldehyde with pFrmR formaldehyde sensing promoter (BBa_K4813003), while the other part (BBa_K4813005) acted as a positive control, where the expression of dTomato was driven by a strong constitutive promoter (BBa_J23100).
Comparing the colouration of dTomato BBa_K4813000 and tdTomato BBa_K4813001
While the fluorescence protein database indicates that tdTomato exhibits superior fluorescence emission compared to dTomato [2], we don't know if it also applies to their chromoprotein properties. Given that our project focuses on developing a device to monitor formaldehyde levels that can be easily used by the general population without specialized equipment, we are interested in finding a chromoprotein that produces a more pronounced color visible to the naked eye.
To address this, we have generated two composite parts, one expressing the dTomato part (BBa_K4813002) and the other expressing tdTomato (BBa_K4813004), and subsequently compared their colors as observed by the naked eye.
Consequently, we opted to use dTomato for the final assay, as it proved to be more effective in detecting the presence of formaldehyde.
Reference
[1] dTomato at fpbase. FPbase. Accessed 20 June 2023 https://www.fpbase.org/protein/dtomato/
[2] tdTomato at fpbase. FPbase. Accessed 20 June 2023 https://www.fpbase.org/protein/tdtomato/
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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