Team:MADRID UCM HS/Demonstrate
Demonstrate
It works!Introduction
Nowadays, society is getting aware of the environment situation. The climate change is a fact due to the high emissions of polluting gases and the use of petroleum. The STAIN project comes up as an initiative to develop sustainable solutions for the planet. Today, producing inks is highly polluting so, how about creating eco-friendly pigments using synthetic biology?
First results of our inks!, Synthesizing orange color
Our main goal is to produce inks of different colors using Synthetic Biology. Bacteria are our perfect machines for this task, using as a model the metabolic route of flavonoids and a precursor called Naringenin. By assembling a promoter (BBa_K2656007), a ribosomal binding site (BBa_K2656009), the gene for the flavanone3-hydroxylase F3H (BBa_K3255000) and a terminator (BBa_K2656026) in a pARK1 vector (α1), we demonstrated that the modified bacteria are capable of producing a light orange/yellow color in presence of the precursor.
Schematic illustration of the assembly of a transcriptional unit (TU), composed by promoter (P), a ribosomal binding site (RBS), a gene and a terminator (TT) in a vector, using the Golden Gate Assembly strategy. The Golden Gate Assembly strategy enables the assembly of these four DNA fragments in a single tube using restriction enzyme BsaI-HFv2 to digest both the vector and the fragments through the recognition of asymmetric DNA sequences and cleavage outside of these recognition sequences, and the T4 DNA ligase to assemble the TU into the vector.
Our objective has been successfully accomplished. In the figure below we show a picture of the cultures exhibiting a deeper yellow in presence of increasing amounts of precursor, and the spectra of these cultures with the highest absorbance at 400nm at the maximum naringenin concentration.
This experiment evidences that parts BBa_K3255000 and BBa_K3255001 work as expected.
Cultures exhibiting a deeper yellow in presence of increasing amounts of precursor, and the spectra of these cultures with the highest absorbance at 400nm at the maximum naringenin concentration.
