Production of Human Alpha 2b Interferon in E.coli Expression System

[摘要] BackgroundandObjective:Interferonsarepartofthebody#039;sdefensesystemagainstviruses.Theseproteinsaregeneratedinthebody,andthentheytriggerthecellsaroundthemtoproduceinhibitorproteinsagainstvirusreplication.Thepurposeofthisstudywascloningandexpressionofthehumaninterferonalpha-2bgeneinpreplasmicspaceofEscherichiacoli.SubjectsandMethods:Inthisresearch,Alpha2binterferongenewasclonedinapET-26b(+)expressionvector,underthecontrolofT7promoterandpelBperiplasmicsignalpeptidesequenceusingNcoIandXhoIrestrictionenzymes.TheexpressionvectorwastransformedintoEscherichiacolistrainBL21(DE3).CloningofalphainterferongeneconfirmedusingcolonyPCR,digestionandDNAsequencing.GeneexpressionofalphainterferonwasexaminedbySDS-PAGEandDotblotanalysis.AlsothepossibilityofperiplasmictargetingwasdeterminedusingSDS-PAGEanddotblotanalysisat15hrsafterinduction.Results:TheresultsshowedthatAlpha2binterferonwasproducedinbacterialexpressionsystemandtargetedtoperiplasmicspace.Conclusion:Ourstudyshowedthattheproductionofpharmaceuticalrecombinantproteininpreplasmicspaceofbacterialexpressionsystemisfissibleandcanbeustilizedasacheapersourceandmoreefficientthanconventionalexpressionsystems.

[学科分类] 基础医学