Cloning,Expression and Transcriptional Activity Assay of Human EYA Gene Family
- VernacularTitle:人EYA基因家族的克隆、表达及转录活性分析
- Author:
Bin YUAN
;
Zhi-Hong XIONG
;
Li-Hua DING
;
Ju-Qiang HAN
;
Hao ZHANG
;
Zhao-Yun WANG
;
Jie-Zhi LI
;
Qi-Nong YE
;
- Publication Type:Journal Article
- Keywords:
Eyes absent Cloning Gene expression Transcriptional activity
- From:
China Biotechnology
2006;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
The complete coding sequences of Eya gene family was amplified by standard PCR fromhuman tissues or cells cDNA library.The product of PCR was cloned into the eukaryotic expression vector pcDNA3-FLAG,generating pcDNA3-FLAG-Eya1～4.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.
