28039.afmnpv誘導的sl1凋亡細胞中凋亡體的檢測

1、華中師范大學碩士學位論文AfMNPV誘導的Sl1凋亡細胞中凋亡體的檢測姓名：張瑋瑩申請學位級別：碩士專業(yè)：農(nóng)業(yè)昆蟲與害蟲防治指導教師：彭建新20100501AbstractApoptosisisakindofindependent，orderlyandaccordingtosomescheduledprogramofnaturaldeathprocessApoptosisisoneofthemostpopularfieldinlifes

2、cienceManyrelatedstimulicomingfromintracellularorextracellularresultinthereleasingofcytochromeCThenthec”ochromeCquicklycombineswiththeapoptoticproteaseactivatingfactorl(Apaf1)。蘆&P|aⅪPandprocaspase一9，whichformsamultimeric

3、proteincomplex(700kD曲calledapoptosomeTheactivationofCaspase9subsequentlytriggersacascadeofeffectorcas_paseactivation，leadingtoapoptoticcelldeathIt’Sveryimprotantformitochondrialsignalpathwayandembryonicdevelopmentinmamma

4、lInthispapertheexistenceofapoptosomeofinsectshasbeenpreliminaryresearchedVirusiSacommoninductionfacorofapoptosisInthisstudySI1cellswereinducedbyAfMNPVwhichiSakindofbaculovirusDAPIstain，agarosegelelectrophoresis，fluoresce

5、ncequantitative，andWestemBlottinghaveusedtodetectthecharacteristicsofmorphologyandbiochemistrytheactivityofCaspase一9，andtheexistenceofapoptosomerespectivelyTheDAPIstainandagarosegelelectrophoresisindicatedthatcellapoptos

6、ishavehappenedafer8hwhichinducedbyA心心口VCaSp舔e9willbeactivatefirst，oncetheapoptosomehasformedTherefore，weusethespecificfluorescencesubstrate(AcLEHDAFC)andspecificinhibitors(ZLEHDFMK)ofCaspase一9todetecttheactivityofCasDase

7、9TheresultsshowthattheactivityofCaspase一9Wasmeasuredafer8hwhichinducedbyAfMNPVandtheactivityineasedtothemaxat24hInordertodetecttheexistenceofapoptosome，weuseWesternBlottingtoanalyzetheproteincomponentofapoptosomeTheprote

8、insthatcomposethecomplexCanbedetectedafer6hwhichinducedbyAfMNPVandthentheydisappearedat12hTheseaboveresultspreliminaryshowthat，asmammaliancellapoptosis，insectcellapoptosisaloshavetheapoptosomeformationKeywords：apoptosis；