Title: Construction and verification of eukaryotic expression plasmid carrying human E2F6 gene
Abstract: Objective To construct and certificate an eukaryotic expression plasmid carrying human E2F6 gene.Methods E2F6 gene was amplified by RT-PCR with the total RNA of human blood plasma as template.The PCR fragments were cloned into pFLAGCMV10 vector to construct recombinant eukaryotic expression plasmids.293T cells were transfected with the recombinant plasmids.RT-PCR was performed to evaluate the transcription of E2F6 in 293T cells.Western blot was used to detect the expressions of E2F6 in 293T cells.Results The sequence of E2F6 was 100% homology with human E2F6 gene previously registered in GenBank.An interesting band about 35-kD was visible in the result of Western blot,which was consistent with expected size of recombinate protein of E2F6 expressed in 293T cells.Conclusion pFLAGCMV10-E2F6 has been constructed successfully.
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
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