NaClO胁迫条件下单增李斯特菌抗氧化胁迫相关功能基因表达轮廓分析
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- 英文论文题名：Anti-oxidation related genesexpression profile analysis in L.monocytogenes response to sodium hypochlorite stress
- 论文作者：周密 ; 唐毓祎 ; 王旭 ; 潘迎捷 ; 张炜佳 ; 赵勇
- 英文论文作者：Zhou Mi ; Wang Xu ; Tang Yuyi ; Zhang Weijia ; Pan Yingjie ; Zhao Yong ; College of Food Science and Technology ; Shanghai Ocean University ; Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation Shanghai ; Ministry of Agriculture ; Shanghai Engineering Research Center of Aquatic-Product Processing and Preservation
- 年：2016
- 作者机构：上海海洋大学食品学院;上海水产品加工及贮藏工程技术研究中心;农业部水产品贮藏保鲜质量安全风险评估实验室(上海);
- 论文关键词：单增李斯特菌 ; 次氯酸钠 ; 氧化胁迫 ; RT-PCR
- 英文论文关键词：listeria monocytogenes ; sodium hypochlorite ; oxidative stress ; RT-PCR
- 会议召开时间：2016-11-09
- 会议录名称：中国食品科学技术学会第十三届年会论文摘要集
- 英文会议录名称：Abstracts of the 13th Annual Meeting of CIFST
- 语种：中文
- 分类号：Q78;Q93
- 学会代码：ZGSP
- 会议名称：中国食品科学技术学会第十三届年会
- 会议地点：中国北京
- 主办单位：中国食品科学技术学会
- 学会名称：中国食品科学技术学会
- 页数：2
- 文件大小：98k
- 原文格式：O
- 会议级别：全国
摘要
为了探究单增李斯特菌氧化还原相关基因抗次氯酸钠胁迫机制,我们在比较不同胁迫时间下过氧化氢酶(CAT)及超氧化物歧化酶(SOD)的酶活变化基础上,通过RT-PCR及主成分分析法观察单增李斯特菌氧化还原相关基因不同时间长度次氯酸钠(30mM)胁迫下观察基因表达变化。结果显示,CAT及SOD仅在胁迫前期(15min,30min)有较强的活性。在NaClO胁迫前期(10min,20min),基因lmo1604,ohrR,perR及sod表达活跃。而在之后的阶段调节细胞氧化还原平衡和蛋白损伤修复的基因表达更为活跃。在这些基因中,grx及lmo1433在30min和40min表达水平显著高于其他基因,而msrA,mrsB及trxA在50 min和60 min时有更活跃的表达。这说明单增李斯特菌主要抵御次氯酸钠的手段(解毒作用和修复作用)并不是同时作用的。此外,结果中硫氧还原蛋白系统相关基因(msrA,mrsB及trxA)只在胁迫后期有较高的表达量,且在额外加入蛋氨酸后细菌的存活率和毒性均显著增强。说明次氯酸钠胁迫造成的蛋氨酸饥饿及细菌毒性下降可能是引起trxA表达滞后的主要原因。
Listeria monocytogenes is a Gram-positive,food-borne pathogenic bacteria which can adapt to serious environment condition.However,the mechanism of NaClO-activated protection byanti-oxidative related genes is not clear.Here,to explore this problem,several genes involved in redox regulation were studied in L.monocytogenes under NaClO stress.Catalase(CAT) and superoxide dismutase(SOD) activities were determined by using the catalase assay kit and the total superoxide dismutase assay kit with NBT,respectively.Gene expression profiling of the pathogen has been investigated using RT-qPCR to understand how L.monocytogenes responds to the condition encountered median lethal concentration NaClO(30mM) stress at differentstresstime.ANOVA,principal component analysis(PCA) were used for further analysis.As the data indicated,catalaseandSOD activities only present a high activity at earlyphase(15minand 30min) andgenetrxA,ohrR and perR were more active during the infection.At the early phase of NaClO stress,genes(Zmo1604,ohrR,perR and sod) known to play a role in NaClO detoxicated presented higher expression levels.However,some redox homeostasis regulate and protein repair related genes performed outstandingly during the subsequent phase.In these genes,glutathione redox system related genes(grx,lmo1433) effected firstly,and thioredoxin system related genes(msrA,mrsB,trxA) played a much more significant role in lastphase.Results of this study showed that the main methods(detoxification and repair effect) of resisting NaClO stress in L.monocytogenes were not acting simultaneously.Thioredoxin system related genes only presenting a high activity in lastphase.And the survival rate and toxicity of bacteria are increased after methionine addition.This indicates that NaOCl stress may cause methionine auxotrophy and virulence decrease.
Listeria monocytogenes is a Gram-positive,food-borne pathogenic bacteria which can adapt to serious environment condition.However,the mechanism of NaClO-activated protection byanti-oxidative related genes is not clear.Here,to explore this problem,several genes involved in redox regulation were studied in L.monocytogenes under NaClO stress.Catalase(CAT) and superoxide dismutase(SOD) activities were determined by using the catalase assay kit and the total superoxide dismutase assay kit with NBT,respectively.Gene expression profiling of the pathogen has been investigated using RT-qPCR to understand how L.monocytogenes responds to the condition encountered median lethal concentration NaClO(30mM) stress at differentstresstime.ANOVA,principal component analysis(PCA) were used for further analysis.As the data indicated,catalaseandSOD activities only present a high activity at earlyphase(15minand 30min) andgenetrxA,ohrR and perR were more active during the infection.At the early phase of NaClO stress,genes(Zmo1604,ohrR,perR and sod) known to play a role in NaClO detoxicated presented higher expression levels.However,some redox homeostasis regulate and protein repair related genes performed outstandingly during the subsequent phase.In these genes,glutathione redox system related genes(grx,lmo1433) effected firstly,and thioredoxin system related genes(msrA,mrsB,trxA) played a much more significant role in lastphase.Results of this study showed that the main methods(detoxification and repair effect) of resisting NaClO stress in L.monocytogenes were not acting simultaneously.Thioredoxin system related genes only presenting a high activity in lastphase.And the survival rate and toxicity of bacteria are increased after methionine addition.This indicates that NaOCl stress may cause methionine auxotrophy and virulence decrease.
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