Holmes Ashleigh, Pritchard Leighton, Hedley Peter, Morris Jenny, McAteer Sean P, Gally David L, Holden Nicola J
Cellular and Molecular Sciences, James Hutton Institute, Dundee DD2 5DA, United Kingdom.
Strathclyde Institute for Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, United Kingdom.
Data Brief. 2020 May 28;31:105769. doi: 10.1016/j.dib.2020.105769. eCollection 2020 Aug.
A high-throughput positive-selection approach was taken to generate a dataset of Shigatoxigenic (STEC) O157:H7 genes enriched in adherence to plant tissue. The approach generates a differential dataset based on BAC clones enriched in the output, after adherence, compared to the inoculum used as the input. A BAC clone library derived from STEC isolate 'Sakai' was used since this isolate is associated with a very large-scale outbreak of human disease from consumption of contaminated fresh produce; white radish sprouts. Spinach was used for the screen since it is associated with STEC outbreaks, and the roots provide a suitable site for bacterial colonisation. Four successive of rounds of Sakai BAC clone selection and amplification were applied for spinach root adherence, in parallel to a non-plant control. Genomic DNA was obtained from a total of 7.17 × 10 cfu/ml of bacteria from the plant treatment and 1.13 × 10 cfu/ml of bacteria from the no-plant control. Relative gene abundance of the output compared to the input pools was obtained using an established DNA microarray chip for STEC. The dataset enables screening for genes enriched under the treatment condition and informs on genes that may play a role in plant-microbe interactions.
采用高通量正选择方法生成了一个富含与植物组织黏附相关基因的产志贺毒素大肠杆菌(STEC)O157:H7基因数据集。该方法基于与用作输入的接种物相比,在黏附后输出中富集的BAC克隆生成一个差异数据集。使用了源自STEC分离株“阪崎”的BAC克隆文库,因为该分离株与因食用受污染的新鲜农产品(白萝卜芽苗菜)引发的大规模人类疾病暴发有关;选用菠菜进行筛选是因为它与STEC暴发有关,且其根部为细菌定殖提供了合适的位点。对菠菜根部黏附进行了四轮连续的阪崎BAC克隆选择和扩增,并与非植物对照平行进行。从植物处理组的细菌中总共获得了7.17×10 cfu/ml的基因组DNA,从无植物对照组的细菌中获得了1.13×10 cfu/ml的基因组DNA。使用已建立的STEC DNA微阵列芯片获得输出与输入文库相比的相对基因丰度。该数据集能够筛选在处理条件下富集的基因,并揭示可能在植物-微生物相互作用中起作用的基因。