牛MYOZ2基因启动子克隆及活性分析

doi:10.11843/j.issn.0366-6964.2016.04.003

Abstract

Abstract:

The Myozenin2 (MYOZ2) gene promoter of bovine was cloned and sequenced to analyze the active region，which will provide theoretical basis for function determination and expression regulation of MYOZ2 gene.Applying 5′-rapid amplification of cDNA end analysis (RACE)，we determined the transcription initiation site of MYOZ2 gene.Using genome DNA as the template，the promoter of MYOZ2 gene was obtained by PCR.Online software was used to analyze putative transcription factor binding sites.Primer were redesigned according to the results of online software analysis，seven promoter fragments in different length were obtained by unidirectional deletion and cloned into luciferase reporter gene expression vectors.Then，the vectors were transfected into C2C12 cells，their expression activity were measured using the dual reporter assay.The promoter sequence of 2 065 bp of MYOZ2 gene in bovine was obtained and the transcription initiation site of MYOZ2 gene was determined.Luciferase assays indicated that the transcriptional activity of pMYOZ2-84/+125 was significantly higher than pGL3-Basic (P＜0.01)，and the transcriptional activity of pMYOZ2-683/+125 also was significantly higher than pMYOZ2-263/+125 (P＜0.01).The core functional promoter of bovine MYOZ2 was located in the region -84/+125 relative to the TSS，and the transcriptional factors such as MEF2，SRF，MyoD and YY1 might be involved in the transcriptional regulation of MYOZ2 gene.

CLC Number:

WANG Ming-ming，ZHAO Zhi-dong，LI An-ning，ZHANG Ya-ran，DUAN Mei-yan，LI Shi-jun，WU Sen，WANG Xiao-yu，ZAN Lin-sen. Cloning and Activity Analysis of Bovine MYOZ2 Gene Promoter[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(4): 652-660.

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Cloning and Activity Analysis of Bovine MYOZ2 Gene Promoter

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