【摘要】：蜂房哈夫尼菌是一種條件致病菌,可導(dǎo)致海產(chǎn)品、肉制品、乳制品、果蔬等食品的腐敗變質(zhì)。為了進一步確定群體感應(yīng)機制在食品腐敗中的作用,并針對群體感應(yīng)系統(tǒng)為食品防腐保鮮提供新策略,研究主要采用熒光定量PCR技術(shù),研究了環(huán)境因素對蜂房哈夫尼菌群體感應(yīng)信號分子基因luxI和luxR相對表達量的影響。為了準確研究luxI和luxR基因表達量的差異,本文首先研究了16S rRNA、23S rRNA、recA和gapA等4種候選內(nèi)參基因在不同實驗條件下的表達情況,發(fā)現(xiàn)它們的表達豐度由高到低依次為16S rRNA23S rRNArecAgapA。使用NormFinder和Best Keeper程序分析后,最終確認16S rRNA表達最為穩(wěn)定,并作為內(nèi)參基因用于后續(xù)實驗。研究了不同氯化鈉濃度、pH及溫度對luxI和lux R基因相對表達量的影響。結(jié)果表明,隨著氯化鈉濃度的增加、pH值的上升及溫度的升高,兩種基因的相對表達量同步變化。當(dāng)氯化鈉濃度為1%、pH為7.5或溫度為10℃和25℃時,luxI和luxR基因的表達量較高;而培養(yǎng)條件在氯化鈉濃度3%以上、pH不為7.5或溫度在4℃以下或37℃以上時,luxI和luxR基因的表達受到抑制。
[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，

本文編號：2261423