S100: Non-traditional host characterization and metabolic engineering pipeline as demonstrated with Synechococcus elongatus PCC 7942

Synthetic biology entails the rational design of living systems toward utilitarian ends.  An application of special interest is metabolic engineering, the modification of small organisms to manipulate molecules of particular value.  From a practical standpoint, metabolic engineering requires the selection of a microorganism, understanding and alteration of its genome, quantitation of the result, and iterative improvement.  We present results for the cyanobacteria Synechococcus elongatus PCC 7942, encompassing rapid whole genome sequencing, de novo contig assembly, automated genome annotation, metabolic model reconstruction, and metabolic engineering predictions.  We demonstrate that a relatively simple toolkit and modest upfront investment can enable a rapid transition from a phenotypically interesting organism to a genomically characterized workhorse.  We show that our short-cut methodology for genome sequencing and annotation compares favorably with the publically available S. elongatus data and enables downstream reconstruction of a functional, fully balanced metabolic model.  We use this model to predict genetic interventions, implement these designs in silico with Vector NTI Express Designer, and model the results with the constraint-based analysis toolbox.