简介：ObjectivesToconstructarecombinantplasmidcarryingenhancedgreenfluorescentprotein(EGFP)andhumanvascularendothelialgrowthfactor(VEGF)121geneanddetectitsexpressioninratmesenchymalstemcells(MSCs).MethodsHumanVEGF121cDNAwasamplifiedwithpolymerasechainreaction(PCR)frompCD/hVEGF121andwasinsertedintotheeukaryoticexpressionvectorpEGFPC1.AfterbeingidentifiedwithPCR,doubleenzymedigestionandDNAsequencing.TherecombinantplasmidpEGFP/hVEGF121wastransferredintoratMSCswithlipofectamine.TheexpressionofEGFP/VEGF121fusionproteinweredetectedwithfluorescencemicroscopeandimmunocytochemicalstainingrespectively.ResultsTherecombinantplasmidwasconfirmedwithPCR,doubleenzymedigestionandDNAsequencing.ThefluorescencemicroscopeandimmunocytochemicalstainingresultsshowedthattheEGFPandVEGF121proteinwereexpressedinMSCs48haftertransfection.ConclusionsTherecombinantplasmidcarryingEGFPandhumanVEGFwassuccessfullyconstructedandexpressedpositivelyinratMSCs.ItoffersapromisetoolforfurtherresearchondifferentiationofMSCsandVEGFgenetherapyforischemialcardiovasculardisease.