作　　者： ; ; ; ; ;

机构地区： 南阳理工学院张仲景国医学院

出　　处： 《南阳理工学院学报》 2011年第6期94-98,共5页

摘　　要： 目的:应用全基因组DNA芯片技术分析低盐冷刺激作用下副溶血弧菌基因的转录表达变化。方法:分别采用"低盐持续刺激培养(continuous growth,CTG)"和"中间转入低盐环境培养(shift growth,STG)"。CTG和STG下,分别采用含NaCl浓度为2%和0.66%的MV-5培养基孵育副溶血弧菌,收集菌体,提取RNA,应用全基因组DNA芯片分别比较两个不同的转录表达谱基因变化特点,分析其作用规律。同时,应用实时定量逆转录多聚酶联反应对芯片结果进行验证。结果:和对照组相比,STG实验中,共有205个基因的转录表达发生显著性变化,上调的基因占优势地位;CTG实验中,总计有298个基因的转录表达发生显著性变化,上、下调的基因总体基本趋于平衡状态,没有明显差异。实时定量逆转录多聚酶联反应结果证实其和芯片数据结果有很强的相关性。结论:在低盐这一"胁迫环境"下,副溶血弧菌利用其存在的独特而精细的应对机制,能够顽强的生存下来并繁衍生殖,这一过程中,节能调节处于调控的核心地位。 Objective：To investigate the expression profiles of Vibrio parahaemolyticus under 2% and 0.66% NaCl in vitro by a global-genome DNA microarray.Method：Two different design,＇continuous growth＇（CTG） and＇shift growth＇（STG） design were chose for this experiment.The bacteria were grown in MV-5 containing under 2% and 0.66% NaCl,cell cultures with an OD600 of 1.2 at the middle exponential phase were harvested for RNA isolation.A whole-genome DNA microarray was constructed to analyze expression profiles of V.Parahaemolyticus,mRNA level of each gene was compared with the different profiles.Real-time quantitative RT-PCR was employed to confirm the microarray data.Result：For the STG design,205 genes showed significantly altered expression compared with the control group,up-regulation of the genes were obviously dominant over the down-regulation.While for the CTG design,298 genes appeared obvious changes in expression,the up-regulation genes kept in balance with the down-regulation genes.Real-time RT-PCR validated the reliability of microarray data.Conclusion：The V.parahaemolyticus has the ability to survive and proliferation in response to the environmental stresses of low salt through its extensive gene transcription regulation,and it is the central theorem for the V.parahaemolyticus to ＂save energy＂ during the process.

关 键 词： 副溶血弧菌 芯片 转录调控 氯化钠 节能

领　　域： [生物学]