秀丽隐杆线虫培养特性与保存方法研究

ActaAgriculturaeZhejiangensis19(1):3436,2007,3,,,(,310029):,,16NGM,30%,-80:;;;:Q18:A:1004-1425(2007)01-0034-03CulturecharactersandpreservationmethodofcaenorhabditiselegansPANGLin2hai,DUAi2fang3,LIXiao2jun,HOUYu2hui,GAOXiang(InstituteofPreventiveVeterinaryMedicine,ZhejiangUniversity,Hangzhou310029,China)Abstract:Caenorhabditiselegans,akindoffree2livingnematodes,hasbeenusedasstudyofgeneticfunctionofparasiticnematodesandasexpressionofthepotentialvaccineantigensofimportantparasiticnematodes.Itwasfoundfromtheculturecondition,morphologicalcharacters,andpreservationconditionofC.elegansthatC.elegansgrewwellat16ontheNGMculturemediumandcanbepreservedfortwomonthsintheliquidof30%Glycerolat-80.Keywords:Caenorhabditiselegans;morphologicalcharacters;culture;preservationCaenorhabditiselegans,,,,,,1mm,,[1,2]Brenner1963,,Brenner,530:2005-11-30:(30571395):(1980-),,,,,3,,E2mail:[e-mail];Tel:[phone],NGM,[3][48],,11.11.1.1N2(C.elegans,theBristolstrainN2),1.1.2OP50OP50,©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(1)NGM1000mlNGM:3gNaCl,2.5g,17g,1mol/LK2HPO42KH2PO4(pH=6.0)25ml,975ml,1ml(5mg/ml),1mol/LMgSO41ml,1mol/LCaCl21ml[7](2)LBLB(3)LB+1000mlLB1ml(5mg/ml)1.1.4(1)M915.12gNa2HPO412H2O(6gNa2HPO4),3gKH2PO4,5gNaCl,0.25gMgSO47H2O,(2)S0.1mol/LNaCl,0.05mmol/LK2HPO42KH2PO4(pH=6.0)(3)1g100ml,1%1.21.2.1M9,420min,,M9420min,[8]200lOP50NGMLBLB,16,;200lOP50NGM,4,16,3772h,1.2.210l1%NGM,5min,,1.2.3M95Ep2pendorf,4,;410%10%(S)30%(S)-80,,,OP50NGM,22.1(OP50)NGMLBLB,1672h,NGM,;LB,,;LB,,NGM2.2(OP50)NGM,4,16,3772h,,4,,;16,;37,2.32.3.1,1mm,(12A,B),,,,,,2.3.2,,,(2)2.4,4,30d,,53:©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(400);B.(200)1Fig.1AdultandlarvaofC.elegans2Fig.2MorphologicalcharactersofC.elegans,,-80,,,NGM;10%10%,,NGM;30%-80,,NGM3NGMLBLB,LB,SulstonK[4,5],NGMK,NGM,,NGM,,16,,20d[7]Brenner[3]30%,Sulston[5]10%25,30%-80,,:[1]RiddleD.C.elegans[M].NewYork:ColdSpringHarborLaboratoryPress,1997.220-226.[2],.[J].,2003,20(10):110-112.[3]BrennerS.TheGeneticsofCaenorhabditiselegans[J].Genetics,1974,77(1):71-94.[4]WardS,BurkeDJ,SulstonJE,etal.GenomicorganizationofmajorspermproteingenesandpseudogenesinthenematodeCaenorhabditiselegans[J].JournalMolecularBiology,1988,199(1):1-13.[5]SulstonJ,HodgkinJ.MethodsinthenematodeCaenorhabditiselegans[M].NewYork:ColdSpringHarborLaboratoryPress,1988.587-606.[6]WilliamsPL,DusenberyDB.UsingthenematodeCaenorhabditiseleganstopredictmammalianacutelethalitytometallicsalts[J].ToxicologyandHealth,1988,4(4):469-78.[7]StephenJ,Kenneya,GaryL,etal.PersistenceofEscherichiacoliO157:H7,SalmonellaNewport,andSalmonellaPoonainthegutofafree2livingnematode,Caenorhabditiselegans,andtrans2missiontoprogenyanduninfectednematodes[J].InternationalJournalofFoodMicrobiology,2005,101:227-236.[8]SarwarH,GhazalaH,RandyG.GeneticTransformationofanEn2tomopathogenicNematodebyMicroinjection[J].JournalofInvertebratePathology,1995,66:293-296.()6319(2007)©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.