Multi-monocistronic and multi-variate vectors were designed， built， and tested for the improved production of cyanidin 3-O-glucoside (C3G) in Escherichia coli BL21 (DE3). The synthetic bio-parts were designed in such a way that multiple genes can be assembled using the bio-brick system， and expressed under different promoters in a single vector. The vectors harbor compatible cloning sites， so that the genes can be shuffled from one vector to another in a single step， and assembled into a single vector. The two required genes: anthocyanidin synthase (PhANS) from Petunia hybrida， and cyanidin 3-O-glucosyltransferase (At3GT) from Arabidopsis thaliana， were individually cloned under P， P， and P pr... More
Multi-monocistronic and multi-variate vectors were designed， built， and tested for the improved production of cyanidin 3-O-glucoside (C3G) in Escherichia coli BL21 (DE3). The synthetic bio-parts were designed in such a way that multiple genes can be assembled using the bio-brick system， and expressed under different promoters in a single vector. The vectors harbor compatible cloning sites， so that the genes can be shuffled from one vector to another in a single step， and assembled into a single vector. The two required genes: anthocyanidin synthase (PhANS) from Petunia hybrida， and cyanidin 3-O-glucosyltransferase (At3GT) from Arabidopsis thaliana， were individually cloned under P， P， and P promoters. Both PhANS and At3GT were shuffled back and forth， so as to generate a combinatorial system for C3G production. The constructed systems were further coupled with the genes for UDP-D-glucose synthesis， all cloned in a multi-monocistronic fashion under P. Finally， the production of C3G was checked and confirmed using the modified M9 media， and analyzed through various chromatography and spectrometric analyses.
