非肌肉肌球蛋白ⅱ型重链a蛋白在prrsv感染细胞中的作用

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1、非肌肉肌球蛋白Ⅱ型重链A蛋白在PRRSV感染细胞中的作用PRA蛋白能与Mab2-5G2特异性结合，并且找到了比较准确的结合位点。通过间接免疫荧光实验证明了PRA蛋白能够与Marc-145细胞的细胞膜特异性结合。更重要的是，通过病毒中和实验，荧光病毒中和实验，荧光定量PCR实验证明了PRA蛋白能够在一定程度上阻断PRRSV对Marc-145细胞的感染。将重组质粒pEGFP-N1-PRA转染至Marc-145细胞，与未转染的Marc-145细胞相比，PRRSV感染转染pEGFP-N1-PRA的Marc-145细

2、胞的能力降低了50%。另外，还证明了NMHCII-A的阻断剂(R)-(+)-Blebbistatin对Marc-145细胞的胞质分裂没有影响，但对PRRSV感染Marc-145细胞有阻断作用。这些实验结果充分证明了PRA蛋白在PRRSV感染易感细胞过程中的作用，为进一步研究PRRSV感染细胞的分子机制提供了一定的依据，也为进一步探求PRRS的防控提供了新的思路。关键词：PRRSV；非肌肉肌球蛋白II型重链A蛋白；Marc-145细胞；阻断作用2山东农业大学硕士学位论文Functionofnon-muscle

3、myosinheavychainII-AproteininporcinereproductiveandrespiratorysyndromevirusinfectionofMarc-145cellsAbstractPorcinereproductiveandrespiratorysyndrome(PRRS)isadiseasecausedbyPRRSvirus(PRRSV)whichbelongstothefamilyArteriviridae.PRRSmainlycausesreproductivedis

4、order,includingpremature,abortion,fetaldeath,mummyandporcinerespiratorysyndromeatvarietyofageswithhighmortality.Atpresent,PRRShasprevailedinthemainworldwhilepig-raisingcountryanddistricts.NowadaysthestudytopathogenicandimmunologicmechanismofPRRSVisnotclear

5、.ThereisalsonoproperwayforcontrolingofPRRSVinfection.PRRSVhasahighlyspecificcelltropism,bothinPAMandinthemonkeykidney-derivedcelllines;thevirusentersthroughamechanismofreceptor-mediatedendocytosis.Untilnow,twoPRRSVreceptors(CD163andVimentin)havebeendescrib

6、edonMarc-145cells.CD163isacellularproteininthescavengerreceptorcysteine-richsuperfamily,functionedasinteractingwithPRRSVandmaydeterminethereplicationlevelsofPRRSV.Vimentinisanintermediate-filamentprotein,whichiswidelyexpressedinvariouscelltypes.Anti-viment

7、inAbsblockPRRSVinfectionofMarc-145cells.WhensimianvimentinwasdeliveredtoBHK-21cellsandCRFKcells,theybecamesusceptibletoPRRSV.VimentinisalsothoughttobeinvolvedinthereplicationandtransportationofPRRSVinsidethecellbyformingacomplexwiththeothercomponentsofthei

8、ntermediatefilaments.AninternalimageMab2-5G2whichwasgeneratedagainstidiotypicantibodiesspecificforGP5antigenofPRRSVidentifiedasoluableproteinpreparedfromMarc-145cells.Theproteinwasidentifiedtobenon-musclemyos