Investigating the impact of heterologous expression of the Micrococcus luteus FabH on global transcription in Escherichia coli

Investigation of whole genome gene expression level changes in a E. coli fatty acid overproducing strain with or without heterologous expression of the M. luteus FabH. The strain expressing M. luteus FabH produces more methyl ketones. This study will be further described in Goh, E.B., E.E.K. Baidoo, J.D. Keasling, and H.R. Beller. Engineering of bacterial methyl ketone synthesis for biofuels. A 11 microarray study using total RNA recovered from six separate control cultures of Escherichia coli K-12 DH1 fatty acid overproducing strain with empty vector and five separate cultures of test strain, Escherichia coli K-12 DH1 fatty acid overproducing strain with vector overexpressing M. luteus FabH. Each chip measures the expression level of 4,254 genes from Escherichia coli K-12 with eight 60-mer probe pairs (PM/MM) per gene, with 2-fold technical redundancy.

本研究针对携带或不携带异源表达藤黄微球菌（M. luteus）FabH的大肠杆菌（Escherichia coli, E. coli）脂肪酸高产菌株的全基因组基因表达水平变化展开探究。其中，表达藤黄微球菌FabH的菌株可产生更多甲基酮。本研究的详细内容将刊载于Goh、E.B.、E.E.K. Baidoo、J.D. Keasling及H.R. Beller发表的论文《用于生物燃料的细菌甲基酮合成工程》（Engineering of bacterial methyl ketone synthesis for biofuels）。
本研究为一项包含11个微阵列（microarray）的实验，所用总RNA提取自6组独立的对照培养物（携带空载体的大肠杆菌K-12 DH1脂肪酸高产菌株），以及5组独立的测试菌株培养物（携带过表达藤黄微球菌FabH载体的大肠杆菌K-12 DH1脂肪酸高产菌株）。每张微阵列芯片可检测大肠杆菌K-12的4254个基因的表达水平，每个基因对应8条60聚体探针对（PM/MM），并具备2倍的技术重复冗余。