Cyan-magenta-yellow Research Articles

Randomized BioBrick Assembly: A Novel DNA Assembly Method for Randomizing and Optimizing Genetic Circuits and Metabolic Pathways

The optimization of genetic circuits and metabolic pathways often involves constructing various iterations of the same construct or using directed evolution to achieve the desired function. Alternatively, a method that randomizes individual parts in the same assembly reaction could be used for optimization by allowing for the ability to screen large numbers of individual clones expressing randomized circuits or pathways for optimal function. Here we describe a new assembly method to randomize genetic circuits and metabolic pathways from modular DNA fragments derived from PCR-amplified BioBricks. As a proof-of-principle for this method, we successfully assembled CMY (Cyan-Magenta-Yellow) three-gene circuits using Gibson Assembly that express CFP, RFP, and YFP with independently randomized promoters, ribosome binding sites, transcriptional terminators, and all parts randomized simultaneously. Sequencing results from 24 CMY circuits with various parts randomized show that 20/24 circuits are distinct and expression varies over a 200-fold range above background levels. We then adapted this method to randomize the same parts with enzyme coding sequences from the lycopene biosynthesis pathway instead of fluorescent proteins, designed to independently express each enzyme in the pathway from a different promoter. Lycopene production is improved using this randomization method by about 30% relative to the highest polycistronic-expressing pathway. These results demonstrate the potential of generating nearly 20,000 unique circuit or pathway combinations when three parts are permutated at each position in a three-gene circuit or pathway, and the methodology can likely be adapted to other circuits and pathways to maximize products of interest.

High-Throughput Screening of Small Molecule Ligands Targeted to Live Bacteria Surface

The discovery of small molecule ligands targeted to the surface of live pathogenic bacteria would enable an entirely new class of antibiotics. We report the development and validation of a microarray-based high-throughput screening platform for bacteria that exploits 300 μm diameter chemical spots in a 1 in. × 3 in. nanolayered glass slide format. Using 24 model compounds and 4 different bacterial strains, we optimized the screening technology, including fluorophore-based optical deconvolution for automated scoring of affinity and cyan-magenta-yellow-key (CMYK) color-coding for scoring of both affinity and specificity. The latter provides a lossless, one-dimensional view of multidimensional data. By linking in silico analysis with cell binding affinity and specificity, we could also begin to identify the physicochemical factors that affect ligand performance. The technology we describe could form the foundation for developing new classes of antibiotics.

Donor–acceptor polymer electrochromes with cyan color: Effect of alkyl chain length on doping processes

A new series of donor–acceptor–donor (D–A–D) type compounds consisting of dialkyl substituted 3,4-propylenedioxythiophene and benzothiadiazole units were synthesized and polymerized chemically and electrochemically to investigate the effect of alkyl chain length on the doping process of the conjugated polymers. It was found that the alkyl chain length plays a key role on both doping processes and the solubility of the polymer. It is also noteworthy that this new series of D–A–D electrochromes transmits or reflects the cyan color of the Cyan–Magenta–Yellow (CMY) color space in the neutral state which can be switched to transmissive grey when oxidized.

Mixed Color Sequential Technique for Reducing Color Breakup and Motion Blur Effects

This paper proposes a mixed color sequential (MCS) algorithm with high contrast enhancement technique in RGB light-emitting diode (LED) backlight display. Owing to synchronous control of liquid crystal display (LCD) and LED panels, high quality image with suppressed color breakup (CBU) and motion blur effects is achieved by our novel color sequential technique. Importantly, MCS algorithm is useful for color filter-less optical compensated bend (OCB) panel display for alleviating CBU and motion blur effects. Furthermore, high contrast image is also presented on LCD panel because of mixed red-green-blue (RGB) and cyan-magenta-yellow (CMY) backlights with optimum power consumption. In other words, MCS algorithm with high contrast enhancement technique can have the better performance compared with other field sequential color techniques. Experimental results demonstrate by an actual RGB backlight module for 32-in 1366*768 LCD panel the improvement of CBU and motion blur effects.

Fabrication of Functionally Graded Materials Via Inkjet Color Printing

A new method for fabricating functionally graded materials (FGMs) via inkjet color printing is reported in this paper. Al2O3 and ZrO2 aqueous suspensions were stabilized electrostatically and placed in different color reservoirs in inkjet cartridges. The volume and composition of the suspensions printed in droplets at a small area were controlled by the inkjet cyan–magenta–yellow–black color printing principle. The analysis of energy‐dispersive spectrometry shows that with multi‐layer printing, the composition profile of the printed FGM is consistent with the designed profile. The new method shows the potential for fabricating FGMs with arbitrarily designed three‐dimensional composition profiles.