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Abstract Myogenin (MyoG) plays a central control role in the process of muscle differentiation. The aim of this study is to research the activity efficiency and tissue specificity of bovine(Bos) myogenin (MyoG) gene promoter. In order to construct a eukaryotic expression vector pDsRed-BosMyoG, promoter fragment of bovine MyoG was obtained by PCR, and pDsRed2 was used as an exogenous vector in which red fluorescence was as a target gene. Then pDsRed-BosMyoG was transfected into a sheep(Ovis aries) skeletal muscle satellite cells and fibroblasts using lipofectamine. Red fluorescence was detected by RT-PCR and Western blot to study activity efficiency and tissue specificity of bovine MyoG-promoter. The results showed that red fluorescence could be observed in transgenic skeletal muscle satellite cells under microscope but not in transgenic fibroblasts. In mRNA level, red fluorescence mRNA expression value in transgenic muscle cells was significant higher than that in the transgenic fibroblasts(P＜0.01). Similarly, Western blot result showed that red fluorescence protein of transgenic skeletal muscle satellite cells was high, but there was not red fluorescence protein of transgenic fibroblasts. The conclusion was that bovine MyoG-promoter can specifically promote the expression of exogenous genes in skeletal muscle cells, and was an effective muscle cells-specific promoter. Cloning bovine MyoG-promoter and exploring the activity of promoter region, was good to reveal a key expression sites of MyoG-promoter and the underlying regulation mechanism of muscle development, and to provide experimental evidences of improvement of meat quality in livestock.
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Received: 09 June 2013
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