Construction of recombinant plasmid pEGFP-anxA5 and its expression in Hela cells

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Objective:To construct the recombinant plasmid pEGFP-anxA5 capable be expressed in mammalian cells.Methods:Annexin A5 gene containing Xho I and Barn HI endoenzyme sites were obtained by PCR.Double enzyme digestion was conducted for green fluorescent protein vector pEGFP-N1 and PCR product of annexin A5 gene.Both fragments were connected by T4 ligase and then transferred to DH5α.After the recombinant plasmid was transfected into Hela cells,the expression of annexin 45 was observed with fluorescent microscope and im- munocytoehemistry staining.Results:Enzyme digestion,PCR and sequencing confirmed the successful construc- tion of the recombinant plasmid,which was highly expressed in Hela cells.Conclusion:The recombinant plasmid pEGFP-anxA5 is successfully constructed and expressed in the uterine cervical cell line Hela cells,which may fa- cilitate the study of annexin A5 function in cervical carcinoma and cervical carcinoma cell lines.

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