O1->O37 serogroup conversion; KanR selected

These experiments were performed to show a serogroup conversion in Vibrio cholerae from O1 to O139. For this purpose, V. cholerae O1 WT = A1552 was grown on crab shell fragments to induce natural competence for transformation. Purified DNA (2 ug each) from strain VC73-orf6/7-Kan-A was added after 24h and the cells grown further for 24h. The VC73-orf6/7-Kan-A strain is a ATCC25873 derivative (both O37 serogroup) which harbors a Kanamycin cassette in the O37 region (as part of the operon between orf6 and orf7 w/o own promotor) for better selection. Transformants were selected on LB+Kan plates. Three clones were selected from each experiment and analyzed by microarray hybridization (BioPrime. Array CGH Genomic Labeling from Invitrogen). Two microarray replicates were done per clone. Comparison of A1552 versus VC73-orf6/7-Kan-A is shown as control. A genotyping experiment design type classifies an individual or group of individuals on the basis of alleles, haplotypes, SNP's. Keywords: genotyping_design Computed

本实验旨在实现霍乱弧菌（Vibrio cholerae）从O1血清群到O139血清群的血清群转换。为此，将O1型野生型霍乱弧菌A1552接种于蟹壳碎片培养基上，以诱导其产生用于转化的自然感受态。24小时后，向体系中加入来自菌株VC73-orf6/7-Kan-A的纯化DNA（每份2μg），继续培养细胞24小时。菌株VC73-orf6/7-Kan-A为ATCC25873的衍生株（二者均属于O37血清群），其O37区域（位于orf6与orf7之间的操纵子内，无自身启动子）携带卡那霉素抗性盒（Kanamycin cassette），以实现更高效的筛选。转化子在添加卡那霉素的LB培养基平板（LB+Kan平板）上筛选获得。每个实验选取3个克隆，通过微阵列杂交（microarray hybridization）进行分析，所用试剂盒为Invitrogen公司的BioPrime阵列比较基因组杂交（CGH）基因组标记试剂盒（BioPrime Array CGH Genomic Labeling）。每个克隆设置2次微阵列重复实验。以A1552与VC73-orf6/7-Kan-A的菌株对比作为对照实验。基因分型实验设计是指基于等位基因、单倍型、单核苷酸多态性（SNP）对单个个体或一组个体进行分类的实验设计类型。关键词：genotyping_design（基因分型设计）、Computed（计算型）