简介:Objective:ToanalyzetypeⅡtopoisomerasegenesinclinicalisolatesoffluoroquinolone-resistantMycoplasmahominis.Methods:EightisolatesofM.hominiscrossresistantto6fluoroquinoloneswereselectedfrom103clinicalstrainsofM.hominisusingabrothmicrodilutionmethod.TypeIItopoisomerasegeneswereamplifiedbyPCRanddirectlysequenced.Nucleotidesequenceswerecomparedtosequencesfromasusceptiblestrain(M.hominisPG2I).Results:MICsofresistantMhisolateswere4-to512-foldhigherthanMICsfromthesusceptiblereferencestrain.SequencecomparisonrevealedaCtoTchangeat113ntingyrAQRDRledtothesubstitutionofSer83byLeucineandnoaminoacidchangeingyrB.AchangeofGtoTat134ntinparCQRDRledtothesubstitutionofSer80byIsoleucineandaGtoAchangeat70ntinparEQRDRledtothesubstitutionofAsparticacidbyAsparagine.Conclusion:TheseresultssuggestthataCtoTchangeatll3ntingyrA,aGtoTchangeat134ntinparCandaGtoAchangeat70ntinpatrEareassociatedwithfluoroquinoloneresistanceofM.hominis.
简介:Objective:ToinvestigatetheexpressionofHPV16mRNAinnormalhumankeratinocytestransfectedwithpSV2-neo/16.Firsthumankeratinocyteswereculturedintheserum-freemediumM154.Second,theplasmidpSV2-neo/16wastransfectedintothehumankeratinocytesusingatransfectingreagent.Third,RT-PCRandSouthernBlottingwereusedtodetecttheexpressionofHPV16mRNAandDNAinthetransfectedkeratinocytes,respectively.Results:TheexpressionofHPV16mRNAwassuccessfullyamplifiedandan110bpwasditectedbyRT-PCR.A7.9kbfragmentwasconfirmedinthetransfectedkeratinocytesbySouthernBlotanalysis.Conclusion:HPV16mRNAandDNAweresuccessfullydetectedinthehumankeratinocytes.