PCR-DGGE 法用于活性污泥系统中微生物群落结构变化解析.docVIP

. . 　　 本文档下载自 HYPERLINK / 文库下载网，内容可能不完整，您可以点击以下网址继续阅读或下载：  HYPERLINK /doc/f2db88e0551810a6f52486e5.html /doc/f2db88e0551810a6f52486e5.html PCR_DGGE法用于活性污泥系统中微生物群落结构变化的解析 第25卷第4期2005年4月 生　　态　　学　　报A CTAECOLOGICASINICA Vol.25,No.4 Apr.,2005 PCR-DGGE法用于活性污泥系统中微生物 群落结构变化的解析 刘新春,吴成强,张　昱,杨　敏,李红岩 (中国科学院生态环境研究中心,环境水质学国家重点实验室,北京　100085) 摘要:应用PCR2DGGE方法,对在相同的操作条件下分别用低温菌和常温菌接种的两套活性污泥系统中的微生物群落结构的动态变化进行了追踪。研究结果表明:由于工艺和操作条件相同,两系统的微生物群落结构的相似性随着运行时间的增加而增加。PCR2DGGE方法可以在一定程度上反应出系统以及操作条件对微生物群落结构变化的影响。关键词:PCR2DGGE方法;活性污泥系统;微生物群落分析 Applicationofpolymerasechainitgelelectro-phoresis(PCR-DGGE)ofmicrobialecologicalcommuns LIUXin2iang,ZHANGYu,YANGMin,LIHong2Yan　 ～847.EcologicaSinica,2005,25(4):842 (StateKeyLaboratoryof EnvironmentalAquaticChemistry,ResearchCenterforEco2EnvironmentalSciences,ChineseAcademyofSciences,Beijing100085,China).Acta Abstract:Populationdynamicsintwoactivatedsludgesystemsinoculatedwithseedsludgecultivatedrespectivelyatalowtemperature((5±1)℃)andnormaltemperature((20±1)℃)werefollowedunderthesameoperationconditionsusingthePCR2/doc/f2db88e0551810a6f52486e5.htmlDGGEmethod. ThegenomicDNAwasextractedfromactivatedsludgeusingphenol2chloroform.PCRamplificationoftheextractedmicrobial16SrRNAgenefragmentswasperformedusingprimersGC341Fand518R.Theresultofagarosegel(118%)electrophoresisshowsthatthePCRproductswereabout230bpinlength.UsingPCRproducts,DGGEwasperformedonaDGGEanalyzerwitha10%(w??v)loadingacrylamide(39:1acrylamide2bisacrylamide)gelundera30%to50%lineargradient.Despitetheofdenaturant.Somebandsappearedinallsamples,whiletheotherswerespecificonlyinspecificsamplesdifferenceofinoculums,however,thebacterialcommunitystructuresinthetwosystemswerequitesimilar. Thesimilarityofcommunitystructuresinthetwosystemsincreasedgraduallyfrom4116%(thefirstday),5012%(thefourthday)upto5913%(thetwentiethday)withtheprolongationofoperationalperiod.ThePCR2DGGEmethodwasa.usefultoolfortheanalysisofpopulationdynamicsinbiologicalsystemsKeywords:activatedsludgesystem;communitystructure;PCR2DGGE文章编号:1/do