新城疫病毒F蛋白七肽重復(fù)序列區(qū)的亞克隆、表達(dá)、純化及結(jié)構(gòu)分析.pdf

1、東北農(nóng)業(yè)大學(xué)碩士學(xué)位論文新城疫病毒F蛋白七肽重復(fù)序列區(qū)的亞克隆、表達(dá)、純化及結(jié)構(gòu)分析姓名：吳庭鶴申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：預(yù)防獸醫(yī)學(xué)指導(dǎo)教師：魏萍2002.5.1ABSTRACTMembranefusionwithintheParmnyxoviridaefamilyofvinIsesismediatedbyasurfaceglycoproteintermedthe‘‘P’orfusionproteinInfectionbyNewcastl

2、eDiseaseVjrus㈣DV)involv笛thefusionofviralandcellularmembraneswi廿lsubsequenttrailsferofviralgeneticmaterialintothecellStudyofthemolecularmechanismofmennbranefusionistargetedforthedevelopmentofnewantivirusesdn瑤sTwoheptadrep

3、eat0IR)regions_mtheN—terminusandCterminUSofmostpararnyxovirusesFproteinincludesofotherenvelopedvirusessuchaSHIVandrespiratorysyneyfialvirus(RSV)，Imvebeenidentified越importanttofusionnamedHR1andHR2n坨twohi眥adrepemregionsc缸f

4、ormcoiledcollhetero_[rimerino_【herenvelopeVi／HSI％，nkformationofeoiledcollheterotrimerCOuldplayakeyroleinvirusmediatedfusionButitiSnotcle口ifNDVhasthesamefusionmechanismasotherenvelopevirI】sesInthisthesis，wehavecloneexpres

5、sedfusionformofFIRlLinkerHR2(HRl2)proteinAndthefusionandfreef01“111ofFIRl，2wempurifiedChemicalcmsslink珊gassayandgelfiltrationconfirmedthatHRl，2canformhomeotrimerTheCircularDichroism(CD)spectroscopyshowedthatHRl衛(wèi)hasahelix

6、characteristic，exhibitingdoubleminimaat208nmand222nmandhaShigIItherlTta]stabilityAll廿|o∞indicatedthattheahelixhomeotrimerformedbyHRI，2ofNDVFproteinisthemoststableformduringfusionprocessandNDVmediatefusionwiththesamemecha

7、tfism∞otherenvelopevirusesDi丘brenttotheotherenvelopedvirusesthereare250anlinoacidsbetweenHRlandHR2Ohoshetal，haverecentlynotedthatNDVFproteinsequencesCOnrainanotherheptadrepeatregionlocatedbetweenHRlandm啦Singlepomtmutatio

8、nsassyshowedthatasingleaminoacidchangeintheHR3sequencesofNDVFproteinc齟alterthestringentrequirementf研HNproteinexpressioninsyncy6LUllformationTo自mlerstudythestructureandfunctionofHR3，wealsohavecloned，expressedthefusionfoIT