Identification of genes induced on nitrate, role of OxyR. Escherichia coli str. K-12 substr. MG1655

The study aimed to identify role of OxyR during growth on different electron acceptors when E. coli are growing anaerobically. Overall design: Wt and OxyR null cells were grown on nitrate and fumarate medium anaerobically. Each condition was done in duplicate. RNA was extracted using Qiagen RNeasy kit after stabilization of RNA with RNA protect bacteria reagent. Hybridization and further processing was done based on Affymetrix protocols on E. coli Genome 2.0 arrays.

本研究旨在明确大肠杆菌（E. coli）厌氧生长时，OxyR在不同电子受体培养条件下所发挥的作用。实验总体设计：将野生型（Wild type，简称Wt）菌株与OxyR缺失（OxyR null）菌株分别接种于硝酸盐培养基与延胡索酸盐培养基中进行厌氧培养，每种实验条件均设置两个生物学重复。先用RNAprotect细菌试剂对细菌RNA进行稳定化处理，再采用凯杰（Qiagen）RNeasy试剂盒提取总RNA。基于Affymetrix实验流程，在大肠杆菌基因组2.0芯片（E. coli Genome 2.0 arrays）上完成芯片杂交及后续处理步骤。