家蠶絲腺生物工廠表達hGM-CSF的研究.pdf

1、蘇州大學碩士學位論文家蠶絲腺生物工廠表達hGMCSF的研究姓名：李曦申請學位級別：碩士專業(yè)：生物化學與分子生物學指導教師：貢成良20090501ExpressionofhGMCSFinSilkwormSilkGlandBioreactorAbstractExpressionofhGMCSFinSilkwormSilkGlandBioreactorAbstractThehumangranulocytemaerophagecolonysti

2、mulatingfactorOaOM—CSF)isanimportantkindoftheglycoproteinthathastheimmunityadjustmentfunctionitplaysthevitalroleinhemopoiesisregulationandtheimmunityadjustm鋤tandhastheimportantclinicalpracticevalueCurrentlyVerietyofthere

3、combinanthGM—CSFexpressionsystemswereintroduced，andInsectexpressionsystemhasitsownadvantagegraduallycometotheforeBombyxmoHisanimportanttraditionaleconomicalinsectkeepingalonghistoryinourcountryandhasacleargeneticbackgrou

4、ndThelarvaofsilkwormhasapairofhigrdydevelopedsilkglands、析tIltheabilitytosynthesizefibroinAtpresentresearchershavemadeitbecomebioreactorandexpressedmanykindsofproteinsHoweveritstillhasmanydisadvantages，includingeffectindu

5、ction，expressionofexogenousgeneandsteadyheredityoftransgenicindividualTransposonexpressionvectorbasedonpiggyBaewasconstructedsuccessfullyinthisthesisThetransgenicvectorcontainsahGMCSFgenecontrolledbyfibroinH—chainpromote

6、randaneomycinresistancegenecontrolledbybaculovirusearlypromoterBmNcellsweretransfected謝mbotlltransposonvectorandhelperplasmidandscreened晰mantibioticsG418AstableexpressiontransformedcelllineWasobtainedELISAassayindicatedt

7、hattheexpressionlevelofhGMCSFinceUlinewasabout153455pg／106cellsInordertointroducethetransposonvectorintoBombyxmoriseveraltransgenicmethodssuchasspermmediatedgenetransferparticledeliverysystemtransfereggprickingandgonadal

8、injectionwereusedAndscreened麗t11GFPandantibioticsG418，combinedidentificationofPCRandSouthernblottransgenicsilkwormswereobtainedTheresultofwesternblotdisplayedthatthemolecularweightofhGMCSFexpressedbytransgcnicsilkwormsil