中国生物工程杂志

The complete coding sequences of Eya gene family was amplified by standard PCR from human tissues or cells cDNA library. The product of PCR was cloned into the eukaryotic expression vector pcDNA3-FLAG, generating pcDNA3-FLAG-Eya1～4.Then human embryo kidney 293T cells were transfected with the recombinant plasmids and the expression of Eya genes were identified by Western blot. Transcriptional assay using a reporter containing myogenin enhance factor indicated that expression of Eya cooperation with Six in 293T cells affected the Myogenin gene expression.The expression vectors of Eya genes were constructed and confirmed by restriction enzyme digestion and DNA sequence analysis. Transcriptional assay using a reporter containing myogenin enhance factor indicated that expression of Eya in coordination with Six in 293T cells stimulated the Myogenin gene expression. This study further confirmed that Eya proteins are transcriptional activator of Six, can improve the activity of myogenin promoter.