Construction of Eukaryotic Expression Vector of Bovine myf6 Gene and Expression of the Gene in Myoblasts

【Objective】 The objective of the study is to construct the eukaryotic expression vector of bovine myf6 gene and analyse the change of gene expression and cell morphology of transfected myoblasts. 【Method】 Eukaryotic expression vector pIRES2-EGFP-myf6 was constructed by insert myf6 in the MCS. Liposome techinigue was used to transfect Luxi cattle myoblasts ,and by G418 selection,a stable transfected cell line was gained. The changes of gene expression of myoblasts were analyzed by western blot and real-time PCR.【Result】 Compared with the control group,the results showed that,after myoblasts transfected by plasmid,the protein and mRNA expression levels of myf6 were both increased (P0.01),muscle creatine kinase gene (P0.01) and myosin light chain gene mRNA expression were both increased (P0.01). Morphology observations indicated that myoblast had fused to myotubes. 【Conclusion】Eukaryotic expression vector pIRES2-EGFP-myf6 can highly express in myoblasts. The results suggest that myf6 gene can promote myoblasts differentiation.