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大肠杆菌 O157:H7 在牛肉提取物与胰蛋白胨大豆肉汤中的差异基因表达。

Differential gene expression of E. coli O157:H7 in ground beef extract compared to tryptic soy broth.

机构信息

U.S. Dept. of Agriculture, Agricultural Research Service, Eastern Regional Research Center, Wyndmoor, PA 19038, USA.

出版信息

J Food Sci. 2011 Jan-Feb;76(1):M79-87. doi: 10.1111/j.1750-3841.2010.01952.x. Epub 2011 Jan 6.

Abstract

E. coli O157:H7 is an important foodborne pathogen, and ground beef is a common vehicle of infection. DNA microarrays have been used for transcriptomic studies of E. coli O157:H7 using laboratory media; however, analysis of gene expression in complex matrices such as food are lacking. This study compared gene expression profiles of E. coli O157:H7 Sakai strain in raw ground beef extract (GBE) and tryptic soy broth (TSB). Total RNA was isolated from GBE and TSB after 2 h of incubation with E. coli O157:H7. Following reverse transcription (RT) of the RNA, labeled cDNA was hybridized to microarrays representing 9608 open reading frames (Operon; Genome Array-Ready Oligo Set) corresponding to 4 genomes of E. coli strains and 3 plasmids. There were 74 up-regulated (genes involved in protein and polysaccharide biosynthesis, transcription factors, membrane transport proteins, and acid shock proteins) and 54 down-regulated (encoding proteins for energy metabolism, biosynthesis of cofactors, transporters of small molecules, and transcription factors and enzymes responsible for protein degradation) genes in E. coli O157:H7 grown in GBE compared to TSB, respectively. Furthermore, compared to incubation in TSB, E. coli O157:H7 incubated in GBE for 2 h showed significantly increased survival when exposed to synthetic gastric fluid, pH 1.5. This study demonstrated that microarray analyses can be performed using complex food matrices, and gene expression of E. coli O157:H7 differs in TSB compared to GBE. The information will be useful for identification of genes that can be employed as potential targets for interventions to control E. coli O157:H7.

摘要

大肠杆菌 O157:H7 是一种重要的食源性致病菌,而绞碎牛肉是常见的感染源。DNA 微阵列已被用于研究大肠杆菌 O157:H7 在实验室培养基中的转录组学,但在食品等复杂基质中分析基因表达的研究还很缺乏。本研究比较了大肠杆菌 O157:H7 菌株 Sakai 在生绞碎牛肉提取物(GBE)和胰蛋白酶大豆肉汤(TSB)中的基因表达谱。将大肠杆菌 O157:H7 分别在 GBE 和 TSB 中孵育 2 小时后,提取总 RNA。对 RNA 进行逆转录(RT)后,将标记的 cDNA 与代表 4 种大肠杆菌菌株和 3 种质粒的 9608 个开放阅读框(Operon;基因组微阵列准备寡核苷酸集)的微阵列杂交。与在 TSB 中相比,在 GBE 中生长的大肠杆菌 O157:H7 有 74 个上调基因(涉及蛋白质和多糖生物合成、转录因子、膜转运蛋白和酸休克蛋白)和 54 个下调基因(编码小分子转运蛋白和转录因子及负责蛋白质降解的酶的能量代谢、辅酶生物合成、)。此外,与在 TSB 中孵育相比,大肠杆菌 O157:H7 在 GBE 中孵育 2 小时后,在暴露于 pH1.5 的人工胃液时,其存活率显著增加。本研究表明,复杂的食品基质可以进行微阵列分析,与 TSB 相比,大肠杆菌 O157:H7 在 GBE 中的基因表达存在差异。这些信息将有助于识别可作为控制大肠杆菌 O157:H7 的潜在干预目标的基因。

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