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.

用于大肠杆菌（Escherichia coli）中目标材料单体生产的组合质粒文库。实验设计：为探究大肠杆菌内目标材料单体的合成，采用D最优实验设计（Design of Experiments, DoE）方法构建了携带目标生物合成途径中各酶促步骤编码基因的组合质粒文库。可变实验因素包括质粒拷贝数（ColE1、p15a、BBR1或SC101复制起点）、启动子强度（Ptrc启动子与PlacUV5启动子）、候选基因筛选，以及质粒内基因与启动子的序列排布。根据实验因素数量及组合空间范围，文库规模设定为2~48个质粒不等。本PDF文件包含组合质粒文库的示意图（图D1至D18），以及用于文库连接酶链式反应（Ligase Chain Reaction, LCR）组装所需的聚合酶链式反应（Polymerase Chain Reaction, PCR）引物序列（表D1）与桥接寡核苷酸序列（表D2）。