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| Study of The Temporal Expression of Listeria monocytogenes Lm319 Virulence Genes |
| 1, 1, 2, 1 |
1.
2. College of Food Science，Agriculture University of Hebei
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Abstract Listeria monocytogenes is an important food-borne pathogen due to its widespread distribution. Consumption of food contaminated with this pathogen can lead to listeriosis. This condition has high rates of morbidity and mortality (25%~30% over all). The organism can endure adverse conditions such as freezing, drying, mild heat and anaerobic conditions. It can also grow at temperatures used in refrigeration (-1.5~4 oC). Due to these characteristics, L. monocytogenes is a great concern for food safety. In this study, the expression timing of Listeria virulence genes in the viral growth cycle was studied to understand the basis for food poisoning caused by pathogens and the effectiveness of food safety control mechanisms. The growth curves for the pathogen were measured at OD600, and 27 of the Lm 319 virulence genes were detected by PCR. Quantitative Real-time PCR (qRT-PCR) was used to analyze 12 batches of 10 virulence genes. The growth curve showed that the first 2 hours of L. monocytogenes Lm319 growth was the lag phase and 2~8 h was the exponential growth phase. During 8~18 h, cell growth entered stationary phase, followed by the declining phase after 18 h. PCR results for 27 virulence genes of L. monocytogenes Lm319 showed that they explicitly contained 23 virulence genes. Quantification by qRT-PCR indicated that 10 virulence genes had similar expression patterns in a 24 h period, with peak expression levels at about 16~18 h in late stationary phase, which thereafter declined but increased again in the death phase. At the same time, it was also found that the expression levels of the 10 virulence genes were significantly different, with invasion associated protein gene (iap), fibronectin-binding protein gene (fbp), hexose phosphate transporter gene (hpt) and host factor for RNA phage Qβ replicase gene (hfq) having the highest expression levels, actin polymerizing protein gene (actA) and hemolysin gene (hlyA) having the lowest expression levels. This study demonstrated that the major virulence genes of L. monocytogenes Lm 319 peaked in expression during the late-stationary phase of cell growth and maintained the high expression levels through the death phase. This conclusion provides a theoretical basis for L. monocytogenes detection and prevention.
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Received: 13 November 2014
Published: 13 April 2015
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