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1、480Adopted:OECDGUIDELINEFORTESTINGOFCHEMICALS23Oct1986"GeneticToxicology:Saccharomycescerevisiae,GeneMutationAssay"1.INTRODUCTORYINFORMATION•Prerequisites–Solid,liquid,vapourorgaseoustestsubstance–Chemicalidentificationoftestsubstance–Purity(impurities)oftestsubstance–Solubilitycharacteristics–M
2、eltingpoint/boilingpoint–pH(whereappropriate)–Vapourpressuredata(ifavailable)•StandarddocumentsTherearenorelevantinternationalstandards.2.METHODA.INTRODUCTIONThisassaymaybeusedtomeasuregenemutationinyeast,aeukaryoticmicro-organism.StrainsofSaccharomycescerevisiaehavebeendevelopedwhichdetectforwa
3、rdorreversemutations(basesubstitutionandframeshift).•DefinitionsBasesubstitutionmutagensareagentswhichcauseabasechangeintheDNA.Inareversemutationassaythischangemayoccuratthesiteoftheoriginalmutation,oratasecondsiteinthegenome.Frameshiftmutagensareagentswhichcausetheadditionordeletionofsingleormu
4、ltiplebasepairsintheDNAmolecule.•PrincipleofthemethodAvarietyofhaploidanddiploidstrainsoftheyeastSaccharomycescerevisiaecanbeusedtomeasuretheproductionofgenemutationsinducedbychemicalagents.UsersofthisTestGuidelineshouldconsultthePreface,inparticularparagraphs3,4,7and8.480page2"GeneticToxicology
5、:Saccharomycescerevisiae,GeneMutationAssay"Forwardmutationsystemsinhaploidstrainssuchasthemeasurementofmutationfromred,adenine-requiringmutants(ade-1,ade-2)todoubleadenine-requiringwhitemutants,andselectivesystems,suchastheinductionofresistancetocanavanineandcycloheximide,havebeenutilized.Themos
6、textensivelyvalidatedreversemutationsysteminvolvestheuseofthehaploidstrainXV185-14Cwhichcarriestheochrenonsensemutationsade2-1,arg4-17,lys1-1andtrp5-48,thatarereversiblebybasesubstitutionmutagensinducingsite-specificmutationsorochresuppressormutations.XV185-14Calsocarriesthehis1-7marker,amissens
7、emutationrevertedmainlybysecondsitemutations,andthemarkerhom3-10whichisrevertedbyframeshiftmutagens.TheonlyextensivelyuseddiploidstrainisD7,whichishomozygousforilv1-92.B.DESCRIPTIONOFTHETESTMETHOD•PreparationsTestsubstanceSo