蜂房哈夫尼菌群体感应信号分子基因表达的研究

发布时间：2018-10-10 10:23

【摘要】：蜂房哈夫尼菌是一种条件致病菌,可导致海产品、肉制品、乳制品、果蔬等食品的腐败变质。为了进一步确定群体感应机制在食品腐败中的作用,并针对群体感应系统为食品防腐保鲜提供新策略,研究主要采用荧光定量PCR技术,研究了环境因素对蜂房哈夫尼菌群体感应信号分子基因luxI和luxR相对表达量的影响。为了准确研究luxI和luxR基因表达量的差异,本文首先研究了16S rRNA、23S rRNA、recA和gapA等4种候选内参基因在不同实验条件下的表达情况,发现它们的表达丰度由高到低依次为16S rRNA23S rRNArecAgapA。使用NormFinder和Best Keeper程序分析后,最终确认16S rRNA表达最为稳定,并作为内参基因用于后续实验。研究了不同氯化钠浓度、pH及温度对luxI和lux R基因相对表达量的影响。结果表明,随着氯化钠浓度的增加、pH值的上升及温度的升高,两种基因的相对表达量同步变化。当氯化钠浓度为1%、pH为7.5或温度为10℃和25℃时,luxI和luxR基因的表达量较高;而培养条件在氯化钠浓度3%以上、pH不为7.5或温度在4℃以下或37℃以上时,luxI和luxR基因的表达受到抑制。
[Abstract]:Hives are conditioned pathogens that cause spoilage in seafood, meat, dairy products, fruits and vegetables. In order to further determine the role of group sensing mechanism in food corruption, and to provide a new strategy for food preservation and preservation by group sensing system, fluorescence quantitative PCR technique was used in this study. The effects of environmental factors on the relative expression of luxI and luxR were studied. In order to accurately study the differences between the expression of luxI and luxR genes, this paper first studied the expression of 16s rRNA,23S rRNA,recA and gapA candidate internal reference genes under different experimental conditions, and found that their expression abundance was 16s rRNA23S rRNArecAgapA. from high to low. The expression of 16s rRNA was confirmed to be the most stable by NormFinder and Best Keeper analysis, and was used as an internal reference gene for further experiments. The effects of pH and temperature on the relative expression of luxI and lux R genes were studied. The results showed that the relative expression of the two genes changed synchronously with the increase of the concentration of sodium chloride and the increase of pH value and temperature. When the concentration of sodium chloride was 7.5 or 10 鈩，

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