一種直接利用含有重組桿狀病毒質粒的大腸桿菌轉染昆蟲細胞的新方法.pdf

1、湖北大學碩士學位論文一種直接利用含有重組桿狀病毒質粒的大腸桿菌轉染昆蟲細胞的新方法姓名：胡貴賓申請學位級別：碩士專業(yè)：遺傳學指導教師：馬立新20090501AbstractThebaculovirusinsectcellexpressionsystem(BES)hasbecomeoneofthemostpopularsystemsfortheexpressionofeukaryoticproteinsinthefieldsofgenet

2、icengineeringHoweveritsextensiveuseislimitedbythetediousmanipulationoflargebaculovirusbacmidDNAsBacteria—mediatedgenedeliveryintomammaliancellsandanimaltissueshasbeendemonstrated，butwhetheritcanbeappliedsuccessfullyinins

3、ectcellsiSunknown—InthisstudyEscherichia—colimediatedegrpgenedeliveryintoinsectcells，includingSpodopterafrugiperda(sf9)andBombyxmori(SmNSWU1)celllines，isdemonstratedMarkedwithenhancedgreenfluorescentprotein，high—titerrec

4、ombinantbaculoviralstockswereharvestedfromcellculturesbysimplecoincubationoftheinsectcellsandEcolistrainscontainingrecombinantbaculovirusbacmidsInternalizedbacteriawerecountedandtheirtitersdetermined，whichindicatedthatre

5、combinantbacmidDNAhadbeentransferredintotheinsectcells，directlymediatedbythehoststrains，andwasfollowedbythepropagationofthebaculovirusesOurresultsconfirmthattheBmNSWU1celllineismoresusceptibletononpathogenicEcolithanareS

6、f9cellsSurprisinglypGB2f2invhlydidnotmarkedlyenhancetheefficiencyofE—colimediatedgenedeliveryintoinsectcellswhichdiffeFSfromtheresultswithmammaliancellsEscherichia—colimediatedgenedeliveryintoinsectcellsishighlyefficient