Protocol to develop a synthetic biology toolkit for the non-model bacterium R. palustris.

Numerous biology tools are developed to work for model organisms, which, however, do not work effectively in non-model organisms. Here, we present a protocol for developing a synthetic biology toolkit for Rhodopseudomonas palustris CGA009, a non-model bacterium with unique metabolic properties. We describe steps for introducing and characterizing biological devices in non-model bacteria, such as the utilization of fluorescence markers and RT-qPCR.

Opening the door to nitrogen fixation in oxygenic phototrophs.

Conveying biological nitrogen fixation (BNF) to photosynthetic species may be the next agricultural revolution, yet poses major engineering challenges. Liu et al. created a diazotrophic strain of a previously non-nitrogen-fixing species, the cyanobacterium Synechocystis sp.

Synthetic Biology Tool Development Advances Predictable Gene Expression in the Metabolically Versatile Soil Bacterium .

Harnessing the unique biochemical capabilities of non-model microorganisms would expand the array of biomanufacturing substrates, process conditions, and products. There are non-model microorganisms that fix nitrogen and carbon dioxide, derive energy from light, catabolize methane and lignin-derived aromatics, are tolerant to physiochemical stresses and harsh environmental conditions, store lipids in large quantities, and produce hydrogen. Model microorganisms often only break down simple sugars and require low stress conditions, but they have been engineered for the sustainable manufacture of numerous products, such as fragrances, pharmaceuticals, cosmetics, surfactants, and specialty chemicals, often by using tools from synthetic biology.