Title: Construction and Expression of Mouse Prx2 Sense or Antisense Eukaryotic Expression Vectors
Abstract: Objective:To construct the sense or antisense mouse Prx2 eukaryotic expression vectors pEGFP-N1-sPrx2 and pEGFP-N1-aPrx2.Methods:Sense and antisense Prx2 full length gene were amplified by RT-PCR from the total RNA of the granulosa cells(GCs) of the mouse ovary,then they were cloned into eukaryotic expression vector pEGFP-N1.After the comformation with PCR, restriction endonuclease digestion and the nucleotide sequencing,the recombinant vectors were transfected into the the GCs which were isolated from the immature mice ovaries transiently through lipofectamine mediation.The expression of Prx2 protein were detected with fluorescent inverted microscope and Western Blot.Results:The sense and antisense Prx2 eukaryotic expression vectors were constructed and verified.The expression of Prx2 protein increased in GCs transfected with pEGFP-N1-sPrx2,while which decreased with pEGFP-N1-aPrx2.Conclusion:The eukaryotic expression vectors pEGFP-N1-sPrx2 and pEGFP-N1-aPrx2 have been constructed successfully.
Publication Year: 2011
Publication Date: 2011-01-01
Language: en
Type: article
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