結(jié)合siRNA技術(shù)的新型多價結(jié)核核酸疫苗的研究.pdf

1、華南理工大學(xué)碩士學(xué)位論文結(jié)合siRNA技術(shù)的新型多價結(jié)核核酸疫苗的研究姓名：程春明申請學(xué)位級別：碩士專業(yè)：發(fā)酵工程指導(dǎo)教師：曹以誠20060524華南理工大學(xué)工學(xué)碩士學(xué)位論文ABSTRACTTuberculosis(TB)isachronicrespiratoryinfectiousdiseasecausedbyMycobacteriumtuberculosis(MTB)Itisestimatedthatonethirdofthewor

2、ldpopulationisinfectedwithMTB，causingover10millionnewTBcasesand3milliondeathsannuallyInChina，thenumberofTBpatientsisthesecondintheworld，approximately400millionpeoplebeinginfected，over5milliongettingsick，and256thousanddyi

3、ngfromTBannuallyTheonlyavailablevaccineagainstTBBCGhasbeenextensivelyevaluatedanddemonstratedavariableprotectiveefficacyrangingfrom0to80％indifferentfieldtrialsFurthermore，duetofollowingissues，suchastheproblemofTBmultidru

4、g—resistant(MDR)strains，CO—infectionwithHIV’andincreasingmobilityofpopulation，theworld—widesituationofTBisdeteriorating，whichhascreatedanurgentneedfornewvaccinestopreventTBA985BandESAT6areconsideredimmunodominantandpromi

5、singvaccinecandidatesasprotectiveantigensforMTBchallengeInthispaper，thegenesrespectivelyencodingA985BandESAT6proteinswereamplifiedbypolymerasechainreaction(PCR)basedongenomeofMTBH37RvstrainThenbygeneSOEingmethod，fusionge

6、neencodingA985BESAT6proteinwaslinkedwiththeIinker(GGIGIAPG)ThefusiongenewasclonedintopVAXItoconstructeukaryoticexpressionplamidpVAXl／AEAfterthat，wemakesomealterationstopVAXl／AEvectorandbypolymerasechainreaction(PCR)，both

7、siMcl一1insertandhiLl2insertwereclonedintopVAXl／AEfromtheconstructedvectorspsiPdqA／Mcl1andpUMVC3hiLl2togettherecombinanteukaryoticexpressionplamidexpressingsiMCLIL、A985B—ESAT6fusedproteinandHill2proteinTheserecombinanteuk

8、aryoticexpressionplamidsrespectivelywerecalledforpVAE、pVAE／siMcl1、pVAE／Hill2andpVAE／siMcll／Hill2Usingsingleanddublerestrictionendonucleasedigestion，PCRandDNAsequencing，itwasconfirmedthattheserecombinanteukaryoticexpressi

9、onplamidshadbeensuccessfullyconstructedTheresultservesasaprototypeofnucleicacidvaccineforfurtherstudiesonitsimmunityeffectivenessagainstMTBInthestudyofTBnucleicacidvaccine，weattemptedtoenhancedtheexpressionofthefusedanti