SYNBIOCHEM - Combinatorial plasmid libraries for material monomer production in Escherichia coli

Combinatorial plasmid libraries for material monomer production in Escherichia coli. - Design of Experiments: To investigate the production of material monomer targets in E. coli, combinatorial plasmid libraries (carrying genes for enzyme steps in target biosynthetic pathways) were designed using a D-optimal design of experiments (DoE) approach. Varied factors include plasmid copy number (ColE1, p15a, BBR1 or SC101 origins of replication), promoter strength (Ptrc or PlacUV5 promoters), candidate gene selection, and sequential ordering of genes/promoters within the plasmid. Library sizes were selected in the range of 2-48 plasmids, depending on the number of factors and combinatorial space. The pdf file contains schematic illustrations of the plasmid libraries (Figures D1-18) along with DNA sequences of the PCR primers (Table D1) and bridging oligosaccharides (Table D2) required for LCR assembly of the libraries. - Learn: Analyses were performed on in vivo screening data for plasmid libraries designed to produce material monomer targets in E. coli. For each plasmid library several analyses were performed in order to assess their predictive capabilities, identify key factors and predict improved constructs. The pdf file collates reports containing the following information: Model fitting; Contrast and regression effects; Model diagnostics; DoE specifications; Predicted best combinations; Predicted best constructs; Design evaluation; Factor power analysis; and Samples relative variance.