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 optimized 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.

[1]

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

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
  COMPATIBLE WITH RFC[1000]