title: Expression of stress and virulence genes in Escherichia coli O157:H7 heat shocked in fresh dairy compost.
abstract: The purpose of this study was to determine the gene expression of Escherichia coli O157:H7 heat shocked in dairy compost. A two-step real-time PCR assay was used to evaluate the expression of stress and virulence genes in E. coli O157:H7 heat shocked in compost at 47.5°C for 10 min. Heat-shocked E. coli O157:H7 in compost was isolated by using an immunomagnetic bead separation method, followed by total RNA extraction, which was then converted to cDNA by using a commercial kit. E. coli O157:H7 heat shocked in broth served as the media control. In compost, heat shock genes (clpB, dnaK, and groEL) and the alternative sigma factor (rpoH) of E. coli O157:H7 were upregulated (P < 0.05), whereas the expression of trehalose synthesis genes did not change. Virulence genes, such as stx1 and fliC, were upregulated, while genes stx2, eaeA, and hlyA were downregulated. In the toxin-antitoxin (TA) system, toxin genes, mazF, hipA, and yafQ were upregulated, whereas among antitoxin genes, only dinJ was upregulated (P < 0.05). In tryptic soy broth, all heat shock genes (rpoH, clpB, dnaK, and groEL) were upregulated (P < 0.05), and most virulence genes (stx1, stx2, hlyA, and fliC) and TA genes (mazF-mazE, hipA-hipB, and yafQ-dinJ and toxin gene chpS) were down-regulated. Our results revealed various gene expression patterns when E. coli O157:H7 inoculated in compost was exposed to a sublethal temperature. Clearly, induction of the heat shock response is one of the important protective mechanisms that prolongs the survival of pathogens during the composting process. In addition, other possible mechanisms (such as the TA system) operating along with heat shock response may be responsible for the extended survival of pathogens in compost.
authors.affiliation: Department of Biological Sciences, Nutrition and Packaging, 228A Life Science Facility, Sciences, Clemson University, South Carolina 29634, USA.
authors.forename: Randhir
authors.initials: R
authors.lastname: Singh
authors.affiliation: Department of Food, Nutrition and Packaging, 228A Life Science Facility, Sciences, Clemson University, South Carolina 29634, USA. xiuping@clemson.edu.
authors.forename: Xiuping
authors.initials: X
authors.lastname: Jiang
mesh_terms: D000818:Animals; D019453:Escherichia coli O157 / Q000235:genetics*; D015870:Gene Expression; D006360:Heat-Shock Proteins / Q000235:genetics*; D018869:Heat-Shock Response / Q000235:genetics*; D006358:Hot Temperature; D018189:Immunomagnetic Separation / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction; D012808:Sigma Factor; D012987:Soil; D012988:Soil Microbiology; D037521:Virulence Factors / Q000235:genetics*
chemical_list: D006360:Heat-Shock Proteins; D012808:Sigma Factor; D012987:Soil; D037521:Virulence Factors