The aims of this proposal are to use DNA arrays and a "systems biology" approach for the analysis and generation of asporogenous, solvent producing strains suitable for continuous or fed-batch bioprocessing, and for studying large cellular programs (sporulation) and phenotypes (solventogenesis and degeneration). Genomic-scale DNA arrays will be used to capture the global picture of cellular transcriptional programs. These studies will be complemented by fermentation and microscopic studies to capture the differentiation process of the various strains. An important aim of both fundamental and practical significance is to determine the gene or genes on the pSOL1-megaplasmid responsible for inhibiting sporulation upon pSOL1 loss. An independent strategy will be also pursued in order to generate asporogenous but solventogenic strains. A third aim is to develop a critical genetic tool for both metabolic engineering (ME) synthesis and functional-genomic analysis, namely that of an efficient recombination system for chromosomal integration and gene deletion based on advanced genetic mechanisms such as the group II intron technology. A fourth aim is to develop strains with new and desirable characteristics in terms of solvent production and efficient bioprocessing, by employing several genetic and ME tools. This project provides unique opportunities for student education and training in the fast growing fields of genomics and ME in an advanced interdisciplinary research environment.
