一種直接利用含有重組桿狀病毒質(zhì)粒的大腸桿菌轉(zhuǎn)染昆蟲細(xì)胞的新方法.pdf

1、湖北大學(xué)碩士學(xué)位論文一種直接利用含有重組桿狀病毒質(zhì)粒的大腸桿菌轉(zhuǎn)染昆蟲細(xì)胞的新方法姓名：胡貴賓申請學(xué)位級別：碩士專業(yè)：遺傳學(xué)指導(dǎo)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