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

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posted on 23.01.2020 by Christopher J. Robinson, Pablo Carbonell, Adrian J. Jervis, Cunyu Yan, Katherine A. Hollywood, Mark S. Dunstan, Andrew Currin, Neil Swainston, Reynard Spiess, Sandra Taylor, Paul Mulherin, Nicholas E. Matthews, Kirk J. Malone, Rosalind Le Feuvre, Philip Shapira, Perdita Barran, Nicholas J. Turner, Jason Micklefield, Rainer Breitling, Eriko Takano, Nigel S. Scrutton
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.

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