PmrA对伤寒沙门菌高渗应激后期基因表达调节初探.doc

PAGE PAGE 1 PmrA对伤寒沙门菌高渗应激后期基因表达调节初探 【摘要】目的：探讨伤寒沙门菌调节因子PmrA在高渗应激后期对基因表达调节的影响。方法：应用自杀质粒介导的同源重组方法制备伤寒沙门菌pmrA基因缺陷变异株；利用伤寒沙门菌全基因组芯片分析技术，比较伤寒沙门菌野生株和pmrA基因缺陷变异株在高渗应激后期的基因表达谱差异，并选择部分差异表达基因进行RT睷CR验证。结果：经PCR及序列分析证实，伤寒沙门菌pmrA基因缺陷变异株制备成功；基因表达谱比较分析结果表明伤寒沙门菌pmrA基因缺陷变异株在高渗应激后期有81个基因表达上调，有22个基因表达下调。结论：PmrA在伤寒沙门菌高渗应激后期的基因表达调节中发挥重要作用。 【关键词】伤寒沙门菌；PmrA；高渗应激；基因芯片 AnalysisofinfluenceofPmrAongeneexpressionregulation ofS.entericaseroverTyphiatlaterstageofhyperosmoticstress GAOYu瞝in,ZHANGHai瞗ang,ZOUXin,DUHong,XIAQiu瞗eng, SHENGXiu瞞ei,XUShun瞘ao,HUANGXin瞲iang (DepartmentofBiochemistry,SchoolofMedicalScienceandLaboratoryMedicine,JiangsuUniversity,ZhenjiangJiangsu212013,China) ［Abstract］Objective:ToexploretheinfluenceofPmrAongeneexpressionregulationofS.entericaseroverTyphiatlaterstageofhyperosmoticstress.Methods：ThepmrAdeletedmutantofS.entericaserovarTyphiwaspreparedbyhomologousrecombinationmediatedbysuicideplasmid.ThehyperosmoticstressenvironmentwassimulatedbyincreasingtheconcentrationofNaClintheLBfrom50mmol/Lto300mmol/Linvitro,andgeneexpressionprofilesofwild瞭ypeandpmrAdeletedmutantofS.entericaserovarTyphiatlaterstageofosmoticstresswereinvestigatedbyS.entericaserovarTyphigeneomemicroarrayanalysis.RT睵CRwasperformedtoprovetheresultsofmicroarrayinsomeselectedgenes.Results：PCRandsequencinganalysisdemonstratedthatthepmrAdeletedmutantofS.entericaserovarTyphihadbeenconstructedsuccessfully;geneexpressionprofilesanalysisrevealedthatexpressionof81genesand22geneswereincreasedanddecreasedrespectivelyinthepmrAmutantatlaterstageofhyperosmoticstress.Conclusion：Theregulator,PmrAofS.entericaseroverTyphi,wasimportantingenesexpressionregulationinresponsetohyperosmoticstress. ［Keywords］S.entericaseroverTyphi;PmrA;hyperosmoticstress;microarray 伤寒沙门菌（SalmonellaentericaserovarTyphi）是沙门菌属中一种人类严重的肠道致病菌，已成为一种重要的原核生物基因表达与信息调控研究的模式菌。目前在沙门菌和埃希菌（Escherichiacoli）中已发现数十种双组分调节系统［1］，PmrAB是其中研究较多的一种，属于经典的二元双组分调节系统［2］。目前研究表明沙门菌PmrAB的活化信号主要有高铁离子（Fe3+）、铝离子（Al3+）、酸性pH（如pH5.5）、钒酸盐［3-5］和低镁离子（Mg2+）［6］,低镁离子的活化是通过PhoPQ系统促进pmrD的表达