一株抗銅鎘細菌的分類鑒定和基因組BAC文庫的構建.pdf

1、、、”’孝十震耄夫學HUAZHONGAGRICULTURALUNIVERSITY碩士學位論文bIASTER’SDEGREEDISSERTATION一標抗銅鎘細菌的分類鑒定和基因組臥c文庫的構津IDENTI丌CATIONOFABACTERIUMRESJS[ANI70CGPPER＆CADMIUMANDCONSTRUCiIONOFGENOMEBACLIBRARY研究牛：導“Ⅱt孥業(yè)：褥園園PANYUANYUAN陳矍莉剮教授ASSOCIATEP

2、ROECItENWENLI糍生物學ⅦCRORIOLOGY囂糊淼黜豁‰。。。船嘰。。中國武漢00E年7iJIAbstractAbacterialstrainNTG01whichwasresistantto45mmol／LCu”and20mmoI／LCd”wasisolatedfromthesoilofacoppermineinDaYecityofHubeiprovinceThehjgllresistanceofthestraintocad

3、miumandcOpperisimportantinstudyingtheresistantmechanismsofbacteriatoheavymetalsExperimentsshowedthattheexpressionofcadmiumresistantgenewasinducedbythecOpperresistantgeneNoplasmidwasfoundinthecellafterplasmidextraction，／n

4、situplasmidfastdetectionandpulsedfieldgelelectrophoresisdeterminationItisassumedthatthegenewastargetedinthechromosomeThisresultwasdifferentfromthepreviousstudiesinwhichtheheavymetalresistantgenewasreportedtobelocatedinpl

5、asmidMorphologicalandphysiologicalidentificationshowedthatNTG01wasGramnegative，shortrodshape，withflagellaaroundthebacteriaVPtestpositiveandMethylredtestnegativeThebacteriumcanuseglucoseascarbonSuOurCCandproducegasesNoH2S

6、producedonTISmediumLysinedecarboxylaseandorthininedecarboxylase，catalaseandgelatinetestarepositiveArgininedccarboxylase，dehydrolase，oxidaseandnraseteStaenegativeSequencingandalignmentof16SrDNAshowedthatitshareda99％homolo

7、goussequencewithEnterobacteraerogenesBasedontheaboveresultsalongwithmorphologicalfeaturesandphysiologicaltraits，thestrainwasidentifiedasEnterobacteraerogenesInthisstudyMICvaluesoftheisolatedstrainto9heavymetalswcredeterm

8、inedandtheM1CvaluesfollowtheorderofAg(000625retool／L)Pb(O9retool／L)Zn(16mmol／L)=CoNi(20retool／L)=CdCr(211retool／L)Cu(45mmol／L)Mn(267mmol／L)ItwasconcludedfromtheMICvalues’thatstrainNTG01ishighlyresistanttomostheavymetals1

9、11egenomiclibrarywasconstructedinthisstudyAccordingtopreviousinvestigationsthesizeofcOpperresistantgeneisgenerally7kbandthefragmentofalltheclonesshouldbefourtimeslagerthanitscontaininggeneThefractionsizeofourgenomiclibra

10、rywasaround40kbSevenhundredcloneswereharvestedandthelibrarycovered99％ofthechromosomeTherefore，thecopperandcadmium—resistantgenecouldbeclonedfromthelibraryintheoryThisstudyattemptedtoclonethecadmiumandcopperresistantgenes