中文摘要：

采用PCR方法从人组织或细胞cDNA文库中扩增Eya基因家族的完整编码序列,将所扩增的基因克隆到带FLAG标签的真核表达载体上,转染人胚肾293T细胞,Western blot鉴定Eya基因家族的表达。检测Eya基因及其与Six基因共转染对MEF3-Luc转录活性的影响。经限制性内切酶分析和DNA序列测定鉴定构建的重组表达载体正确,通过Westernblot实验证明Eya基因家族真核表达载体构建成功。荧光素酶活性测定显示Eya能协同Six激活肌细胞生成素的表达。进一步证实Eya是Six的转录共激活因子,能提高肌细胞增强因子的活性。

英文摘要：

The complete coding sequences of Eya tissues or cells cDNA library. The product of PCR was gene family was amplified by standard PCR fromhuman cloned into the eukaryotic expression vector pcDNA3- FLAG,generating pcDNA3-FLAG-Eyal -4. Thenhuman 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. Eya proteins are transcriptional activator of Six and can improve the activity of myogenin promoter.