Title:

Development of a Synthetic Biology Toolbox for Synechococcus sp. PCC 7002

Authors & affiliations:

Anne M. Ruffing

Sandia National Laboratories, Bioenergy & Defense Technologies, PO Box 5800, Albuquerque, NM

87185

Abstract:

Biofuel and chemical production in cyanobacterial hosts has recently expanded due to the search for renewable alternatives for conventional products of the petroleum industry. Cyanobacteria require minimal resources, utilizing sunlight for energy and carbon dioxide as a carbon source, with some strains even capable of nitrogen fixation. Hence, cyanobacterial-based production offers the promise of reduced feedstock costs and the potential for reduced greenhouse gas emissions compared to traditional fermentation hosts. Despite these potential advantages, cyanobacterial strain development is limited by a
lack of genetic tools and chassis organisms for systems-level optimization. To advanced synthetic biology capabilities for cyanobacteria, this work aims to develop a synthetic biology toolbox (i.e. characterized genetic tools and parts) for the genetic manipulation of Synechococcus sp. PCC 7002. Tools developed for Synechococcus sp. PCC 7002 include fluorescent protein reporters and native promoters with varying expression levels (strong, moderate, and weak) and regulation (constitutive, linear phase, and stationary phase). Additionally, transformation conditions, including cell concentration, length of homologous regions, and the effect of light and temperature on ploidy level, were investigated for the cyanobacterial chassis Synechococcus sp. PCC 7002. The construction and availability of a synthetic biology toolbox is a critical step in advancing cyanobacterial strain development for commercial applications.