预览加载中，请您耐心等待几秒...

鼠伤寒沙门氏菌鞭毛蛋白FliC的原核表达、纯化及其多克隆抗体的制备.docx

预览
1/3
2/3
3/3

在线预览结束，喜欢就下载吧，查找使用更方便

下载提示 文本预览

如果您无法下载资料，请参考说明：

1、部分资料下载需要金币，请确保您的账户上有足够的金币

2、已购买过的文档，再次下载不重复扣费

3、资料包下载后请先用软件解压，在使用对应软件打开

鼠伤寒沙门氏菌鞭毛蛋白FliC的原核表达、纯化及其多克隆抗体的制备 Title:ProkaryoticExpression,Purification,andGenerationofPolyclonalAntibodiesagainstFlagellarProteinFliCofRatHaemophilusSalmonella Abstract: Inthisstudy,weaimedtoachievetheprokaryoticexpressionandpurificationoftheflagellarproteinFliCofRatHaemophilusSalmonellaandsubsequentlygeneratepolyclonalantibodiesagainstit.TheflagellarproteinFliCisacrucialcomponentofSalmonellaflagellaandplaysavitalroleinbacterialmotility.ThisstudywillcontributetoadeeperunderstandingofthepathogenicityandimmunologicalresponseinducedbyRatHaemophilusSalmonella,aidinginthedevelopmentoftargetedantimicrobialstrategies. Keywords:flagellarproteinFliC,prokaryoticexpression,purification,polyclonalantibodies,RatHaemophilusSalmonella Introduction: RatHaemophilusSalmonellaisapathogenicbacteriumthatcausesawiderangeofdiseasesinbothhumansandanimals,includinggastroenteritis,pneumonia,andmeningitis.TheflagellarproteinFliCisaprincipalcomponentofSalmonellaflagella,whichisresponsibleforthemotilityofthebacterium.FliCisknowntobeantigenicandhasbeenimplicatedinhostimmuneresponses.Therefore,theprokaryoticexpression,purification,andgenerationofpolyclonalantibodiesagainstFliCmayprovevaluableforstudyingthepathogenesisanddevelopmentoftargetedtherapeuticinterventions. Methods: 1.Genesynthesisandprokaryoticexpression: ThefliCgeneofRatHaemophilusSalmonellawassynthesizedandclonedintoanexpressionvectorsuitableforprokaryoticexpression.ThetransformedhostE.coliBL21(DE3)wasinducedwithIPTGtoexpressrecombinantFliC. 2.Proteinpurification: Followingexpression,thebacterialcellswerelysed,andFliCwaspurifiedusingaffinitychromatography.Briefly,thelysatewasloadedontoaNi-NTAresincolumnandwashedextensivelytoremoveimpurities.ElutionofFliCwasachievedbyapplyingimidazole,andthepurifiedproteinwasdialyzedtoremoveresidualimidazoleandbufferexchange. 3.SDS-PAGEanalysis: ProteinsampleswererunonSDS-PAGEgelsandvisualizedusingCoomassieBrilliantBluestaining.ThepurityandmolecularweightoftheFliCproteinweredeterminedbycomparingwit