DNA microarray analysis of genes positively regulated by HapR. Vibrio cholerae

Vibrio cholerae strains A1552 and ATN140 (A1552-delta-hapR) were diluted 1:100 from over night cultures and grown aerobically (at 250 rpm) in M9 minimal medium supplemented with vitamins (MEM vitamin solution, BRL) and 0.5% lactate. At OD600 ~ 0.3 2 mM (GlcNAc)6 (Seikagaku America, East Falmouth, MA) was added and the cultures were further shaken for 2 hours. After harvesting the bacteria by centrifugation, the cell pellet was resuspended in Trizol reagent (GIBCO/BRL) and stored at -80 degrees C until further use. The procedure of RNA isolation and the corresponding amplicon microarray experiments was described by K. L. Meibom et al., Proc. Natl. Acad. Sci. U.S.A 101, 2524-9 (2004). Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Keywords: Logical Set Overall design: Computed

将霍乱弧菌（Vibrio cholerae）菌株A1552与ATN140（A1552-ΔhapR）从过夜培养物中以1:100的比例稀释，于添加了MEM维生素溶液（BRL）及0.5%乳酸盐的M9基础培养基中，以250 rpm转速进行有氧振荡培养。当菌液的OD600值约为0.3时，加入2 mM N-乙酰葡糖胺六聚体（(GlcNAc)6，Seikagaku America, East Falmouth, MA），继续振荡培养2小时。通过离心收集菌体后，将细胞沉淀重悬于Trizol试剂（GIBCO/BRL）中，并于-80℃低温保存待用。RNA提取流程及配套的扩增子微阵列实验已由K. L. Meibom等人发表于《美国国家科学院院刊》（Proc. Natl. Acad. Sci. U.S.A）101卷，2524-2529页（2004年）。该微阵列集基于共享的生物学关联进行组织，涵盖物种、肿瘤类型、生物学过程等类别。关键词：逻辑集（Logical Set）。总体设计：计算衍生。