Team:MADRID UCM HS/Parts
Parts
Introduction
During the development of the project we worked to create new parts, adapting the genes we used according to iGEM standardization requirements.
Basic Part
Our part, BBa_K3255000, containing F3H gene codifies for enzyme flavanone3-hydroxylase in the route of anthocyanin synthesis that transforms naringenin, a flavonoid precursor, in dihydrokaempferol. The gene has a plant origin, from Camelia sinensis. Codons had been optimized for E. coli. It contains fusion sites for Golden Gate Assembly.
Composite part
To create a full transcriptional unit (composite part) that contains F3H gene in pARK1 (α 1) vector, we used the promoter BBa_K2656007, the RBS BBa_K2656009 and the Terminator BBa_K2656026 from Valencia UPV team (2018), creating a new composite part, BBa_K3255001. In this process, we validated BBa_K2656007, BBa_K2656009 and BBa_K2656026, as we could show that these parts work in our system (See results)
We have been able to clone genes DRS (dihydroflavonol 4-reductase), ANS (anthocyanidin synthase) and 3GT (anthiocyanin 3-O-glucosylransferase) in vector pDGB1 and their correspondent transcriptional units were cloned in pARK. Cloning was tested by sequencing analysis (see sequencies) but could not be further validated. Measuring enzyme activity doing the corresponding spectra was not informative as the product of these enzymes does not produce a change in color.
