Abstract:

This experiment was conducted to compare the activity of different fragments size of Wagyu MyoG promoter and investigate the potential mechanism. According to the sequence of Bovine myogenin published in GenBank, the primers were designed to amplify the promoter region of Wagyu MyoG by performing PCR. The PCR products were ligated to pMD18TMyoGpro cloning vector. Positive clones were identified by restriction endonuclease and sequencing. Then the Wagyu MyoG promoter dualluciferase reporter gene vector(pGL3MyoGpro) was constructed, and then was transfected into muscle derived stem cells(MDSCs) and fetal fibroblast cells of Bos traurus, and the activity of dualluciferase reporter gene was detected. The activity and the specificity of these promoters during 1662 125 bp were identified in MDSCs. Bioinformatics analysis showed that the promoter fragment contained one TATAbox, fifteen Ebox and may contained binding sites of transcription factors MyoD, MEF2, MEF3, MTBF, TEF1, PRDF, Sp1, SRF, ERE, GRE and Oct1. By comparing the activity of different promoter fragments and analyzing on these transcription factors, it initially indicate that these transcription factors may play an important role in regulation of promoter activity. Cloning and analyzing of the promoter region and function of MyoG lay a foundation for further research on the mechanism of regulation and expression of MyoG in Bos taurus.