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Applied and Environmental Microbiology, November 2008, p. 6859-6866, Vol. 74, No. 22
0099-2240/08/$08.00+0     doi:10.1128/AEM.00356-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Gene Expression Profiling of Listeria monocytogenes Strain F2365 during Growth in Ultrahigh-Temperature-Processed Skim Milk {triangledown} ,{dagger}

Yanhong Liu* and Amy Ream

Microbial Food Safety Research Unit, U.S. Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center, 600 East Mermaid Lane, Wyndmoor, Pennsylvania 19038

Received 12 February 2008/ Accepted 9 September 2008

To study how Listeria monocytogenes survives and grows in ultrahigh-temperature-processed (UHT) skim milk, microarray technology was used to monitor the gene expression profiles of strain F2365 in UHT skim milk. Total RNA was isolated from strain F2365 in UHT skim milk after 24 h of growth at 4°C, labeled with fluorescent dyes, and hybridized to "custom-made" commercial oligonucleotide (35-mers) microarray chips containing the whole genome of L. monocytogenes strain F2365. Compared to L. monocytogenes grown in brain heart infusion (BHI) broth for 24 h at 4°C, 26 genes were upregulated (more-than-twofold increase) in UHT skim milk, whereas 14 genes were downregulated (less-than-twofold decrease). The upregulated genes included genes encoding transport and binding proteins, transcriptional regulators, proteins in amino acid biosynthesis and energy metabolism, protein synthesis, cell division, and hypothetical proteins. The downregulated genes included genes that encode transport and binding proteins, protein synthesis, cellular processes, cell envelope, energy metabolism, a transcriptional regulator, and an unknown protein. The gene expression changes determined by microarray assays were confirmed by real-time reverse transcriptase PCR analyses. Furthermore, cells grown in UHT skim milk displayed the same sensitivity to hydrogen peroxide as cells grown in BHI, demonstrating that the elevated levels of expression of genes encoding manganese transporter complexes in UHT skim milk did not result in changes in the oxidative stress sensitivity. To our knowledge, this report represents a novel study of global transcriptional gene expression profiling of L. monocytogenes in a liquid food.

* Corresponding author. Mailing address: Microbial Food Safety Research Unit, Eastern Regional Research Center, 600 East Mermaid Lane, Wyndmoor, PA 19038. Phone: (215) 233-6587. Fax: (215) 233-6581. E-mail: Yanhong.Liu{at}ars.usda.gov

{triangledown} Published ahead of print on 19 September 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, November 2008, p. 6859-6866, Vol. 74, No. 22
0099-2240/08/$08.00+0     doi:10.1128/AEM.00356-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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