牛磺酸作用下副溶血弧菌基因转录表达变化分析  

Global Gene Expression Profiles of Vibrio parahaemolyticus Induced by Taurocholic Acid

作　　者：杨雷[1,2] 马长德[1] 谢东霞[1] 徐国昌[1] 毛秉豫[1] 周冬生[2] 

机构地区：[1]南阳理工学院张仲景国医学院,河南南阳473004 [2]军事医学科学院微生物流行病学研究所国家重点实验室,北京100071

出　　处：《时珍国医国药》2011年第12期2836-2839,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(No.30871370);国家"863"计划课题(No.2006BAK02A15);国家传染病科技攻关计划(2009ZX10004-103及2008ZX10004-009)

摘　　要：目的利用全基因组DNA芯片技术分析牛磺酸作用下副溶血弧菌基因转录变化概况,界定受牛磺酸直接调控的基因,研究其表达调控变化规律。方法分别采用"牛磺酸持续刺激培养(CTG)"和"中间添加牛磺酸刺激培养(STG)"方案,在副溶血弧菌生长的培养基中添加50 mmol/L牛磺酸,孵育至对数中期OD600=1.2,收集菌体,提取RNA,应用全基因组DNA芯片分别分析比较其和正常对照组的基因转录变化特征,并对比分析两个不同的转录表达谱基因变化特点,界定出可能受牛磺酸直接调控的基因群。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,CTG实验中,总计有255个基因的转录表达发生显著性变化,上调的基因明显占主导优势;STG实验中,共有129个基因的转录表达发生显著性变化,同样以上调的基因占优势地位。综合分析两个表达谱基因转录表达特征表明,44个基因可能受牛磺酸直接影响调控,分别涉及硫代谢、谷胱氨酸代谢、铁离子代谢、核糖体加工合成等。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。结论我们应用DNA芯片技术描绘出了受牛磺酸影响的副溶血弧菌基因转录表达变化概图,并推测,蛤蜊、扇贝等海洋生物体内含量丰富的牛磺酸资源可能为副溶血弧菌的滋生和繁殖提供了重要保障。Objective To dissect expression profiles of Vibrio parahaemolyticus in response to taurocholic acid in vitro by a global-genome DNA microarray,and to define the direct regulatory genes by taurine.Methods Two different design,continuous growth（CTG） and shift growth（STG） design were chosen for this experiment.The bacteria were cultured in MV-5 containing 50mmol/L taurine,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.Then the taurine was defined at the same time.Real-time quantitative RT-PCR was employed to confirm the microarray data.Results For the CTG design,255 genes showed significantly altered expression compared with the control group,while for the STG design,129 genes showed obvious changes in expression.In both profiles,up-regulation of the genes was obviously dominant over the down-regulation,among which the genes were related to the sulphur amino acid,aminoglutaminic acid,iron metabolism and ribosomal proteins：synthesis and modification,displayed a unique pattern.Real-time RT-PCR validated the reliability of microarray data.Conclusion We bring a genome-wide picture of bacterial addition of the taurocholic acid at the transcriptional level,and speculate clams,scallops and other marine resources in the body content of taurine wealth may be an important safeguard for the breeding and reproduction of V.parahaemolyticus.

关 键 词：副溶血弧菌 牛磺酸 芯片 转录调控 

分 类 号：R2-03[医药卫生—中医学]