Gene Expression Changes in Campylobacter jejuni triggered by exposure to Novobiocin. Campylobacter jejuni

In order to investigate the gene experssion changes in Campylobacter jejuni triggered by exposure to Novobiocin. Overall design: DNA microarray was used to identify genes that were differentially expressed in C. jejuni 11168 treated with Novobiocin. For RNA extraction, Campylobacter cells were grown 4 hrs in MH broth to the mid exponential phase (OD600=0.4-0.5) with shaking rates 180rpm at 42°C, and then split into two equal portions, one of which was treated with subinhibitory dose of Novobiocin (256ug/ml, 0.25 MIC) and the other served as a non-treated control, all of these samples were incubated at 42°C for 30 min under microaerobic conditions, RNA samples were extracted from 4 independent treatments and 4 non-treated controls. Then cDNA was synthesized using aminoallyl-dNTP and RNA samples according to the manufacturer’s instruction. After purification, aminoallyl labeled cDNA was fluorescently labeled with Cy-3/Cy-5 mono-Reactive Dye Pack.