Characterization of neutral sites to use Synechocystis sp. PCC 6803 as a photoautotrophic chassisView all posters
IBMC - Instituto de Biologia Molecular e Celular & Faculdade de Ciências, Portugal
Cyanobacteria are promising “low-cost” cell factories since they have minimal nutritional requirements and high metabolic plasticity. The unicellular non-N2-fixing cyanobacterium Synechocystis sp. PCC 6803 is the best studied strain and the vast amount of physiological and molecular data available already allowed the construction of computational genome-scale metabolic models . Moreover, a previous work from our group reported the construction of a Synechocystis chassis prone to receive synthetic devices primarily designed for hydrogen production . Within this work, five neutral sites were identified and characterized to stably integrate ectopic DNA into the chromosome, foreseeing the use of Synechocystis as a photoautotrophic chassis in biotechnological applications. The neutrality of the sites was evaluated by producing and analyzing deletion/insertion mutants, and their functionality was assessed introducing a synthetic device expressing the reporter green fluorescent protein. Moreover, integrative vectors including a multiple cloning site compatible with the BioBrick RFC standard and insulated by transcription terminators were constructed, constituting robust cloning interfaces for synthetic biology purposes. Synechocystis mutants (chassis) ready to receive purpose-built synthetic modules/circuits are also available. References:  Montagud A et al. (2010) BMC Syst Biol 4:156.  Pinto F et al. (2012) Microbiology 158:448-464.