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1、J.Microbiol.Biotechnol.(2005),15(6),1170–1177MonitoringofMicroorganismsAddedintoOil-ContaminatedMicroenvironmentsbyTerminal-RestrictionFragmentLengthPolymorphismAnalysisJUNG,SEONG-YOUNG1,2,JUNG-HYUNLEE1,YOUNG-GYUCHAI2,ANDSANG-JINKIM1*1MicrobiologyLaboratory,
2、KoreaOceanResearch&DevelopmentInstitute,AnsanP.O.Box29,425-600,Korea2DepartmentofBiochemistry,HanyangUniversity,425-791,KoreaReceived:May20,2004Accepted:September19,2004AbatractTerminal-restrictionfragmentlengthpolymorphismrequiresknowledgeofhowlongthestrain
3、ssurviveinthe(T-RFLP)analysiswasusedtomonitorinoculatedoil-degradingtargetenvironment.Tomonitorspecificbacteriaintroducedmicroorganismsduringbioremedialtreatabilitytests.Apairintoenvironments,aswellasindigenousmicrobialgroups,ofuniversalprimers,fluorescently
4、labeled521Fand1392R,microbialstrainsmustbedetectedspecificallyandsensitively.wasemployedtoamplifysmallsubunitrDNAinordertoTraditionally,microorganismshavebeenmonitoredbysimultaneouslydetecttwobacterialstrains,Corynebacteriumcultivationtechniquessuchasplateco
5、unting[7].However,sp.IC10andSphingomonassp.KH3-2,andayeaststrain,becausemostnaturallyoccurringmicroorganismsarenotYarrowialipolytica180.Digestionofthe5'-endfluorescence-cultivatedbystandardculturetechniques,alternativemethodslabeledPCRproductswithHhaIproduce
6、dspecificterminal-mustbeusedtodetectspecificmicroorganisms[13,14].restrictionfragments(T-RFs)of185and442bases,correspondingTherefore,molecularbiologytechniquesthatdonotrequiretoCorynebacteriumsp.IC10andY.lipolytica180,respectively.microorganismcultivationare
7、valuablefordetectingTheenzymeNruIproducedaspecificT-RFof338basesmicroorganismsinnaturalenvironments[1].forSphingomonassp.KH3-2.Thedetectionlimitforoil-Terminal-restrictionfragmentlengthpolymorphism(T-degradingmicroorganismsthatwereinoculatedintonaturalRFLP)a
8、nalysis,basedon16SrDNA,wasdevelopedtoenvironmentswasdeterminedtobe0.01%ofthetotalidentifybacterialspecies[2].Subsequently,ithasbeenmicrobialcount,regardlessofthebackgroundenvironment.usedforstru
