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囊泡法转染重组FSH在巴斯德毕赤酵母中的表达.docx

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囊泡法转染重组FSH在巴斯德毕赤酵母中的表达 Title:ExpressionofRecombinantFSHinPichiapastorisUsingtheSaccharomycescerevisiaeSecretoryPathway Abstract: Follicle-stimulatinghormone(FSH)isakeyreproductivehormonethatplaysavitalroleinfollicledevelopment,maturation,andovulationinfemalesandspermatogenesisinmales.RecombinantFSHproductionisofgreatimportanceforclinicalandresearchpurposes.However,obtaininghighyieldsoffunctionalFSHhasbeenchallenging.Inthisstudy,weutilizedtheSaccharomycescerevisiaesecretorypathwayinPichiapastoristoexpressrecombinantFSHusingthevesicletransfectionmethod.TheaimofthisresearchwastoevaluatetheefficiencyofthisapproachandassessthefunctionalityoftheexpressedFSH. 1.Introduction: Follicle-stimulatinghormone(FSH)isaglycoproteinhormonethatregulatesreproductiveprocessesinbothmalesandfemales.Itstimulatesthegrowthanddevelopmentofovarianfolliclesinfemalesandspermproductioninmales.Duetoitsimportantbiologicalfunctions,recombinantFSHhassignificantpotentialforvariousapplications,includingfertilitytreatment,contraception,andresearchpurposes. 2.ExpressionSystemsforRecombinantFSH: SeveralexpressionsystemshavebeenemployedfortheproductionofrecombinantFSH,includingmammaliancells,insectcells,andbacteria.However,thesesystemshavelimitations,suchaslowyield,highcost,andpost-translationalmodificationdifficulties.Theuseofyeastcells,particularlyPichiapastoris,hasemergedasanattractivealternativeduetoitslow-costfermentation,highcelldensities,andeukaryoticproteinprocessingcapabilities. 3.TheSaccharomycescerevisiaeSecretoryPathway: ThesecretorypathwayofSaccharomycescerevisiaeiswell-characterizedandwidelyusedfortheheterologousexpressionofproteins,includingglycoproteins.Thissysteminvolvestheefficienttranslocationofrecombinantproteinsintotheendoplasmicreticulum(ER)andsubsequentsecretionintotheextracellularmedium.Thevesicletransfectionmethodallowsforthedirecttransferofplasmidsintotheyeastcells,enablingefficientproteinproduction. 4.ConstructionofRecombinantFSHExpressionVectors: ThegeneencodingtheFSHsubunits(αa