The Cloning Of Myf5Gene And Reseaching About The Spatio-temporal Expression Of Myf5Gene In Pig

The Myf5gene, which belong to the family of muscle regulatory factors (MRFs) play a major role in muscle growth and development. It plays a key role in the specification and initiation of myoblast during embryonic myogenesis, and Myf5is expressed in satellite cells and activated satellite cells in adult muscle. The objective of this study was to obtain the coding sequence of Myf5gene in Mashen pig and investigate the level of developmental expression of Myf5gene in porcine skeletal muscles.Firstly, the coding sequence of Myf5gene was obtained by PCR, and two mutation site (C205A、 G735C) were examined. Then analysed the bioinformatic function of Myf5protein, Fourteen O-glycosylation sites and thirty-one phosphorylation sites were predicted by NetoGlyo3.1and NetPhos2.0about amino acids sequence of Myf5protein. Prediction of tertiary structure about Myf5protein was base Helix-loop-Helix.Secondly, the expression patterns of Myf5gene was studied at the levels of mRNA and protein in different tissues of Mashen and Large White pig. The results showed that the Myf5gene just expressed in skeletal muscle, and the relative expression quantity of Myf5gene of Mashen pig in skeletal muscle was greater than large White. However, there was no statistically significant differnces between the muscles in two breeds.Lastly, the expression patterns of Myf5gene was investigated at the levels of mRNA and protein in longissimus dorsi at seven developmental stages (1,30,60,90,120,150, and180-day old) of Mashen and Large White pig. The results showed that the greatest expression of Myf5gene in longissimus dorsi was at one-day old, then at30-day old both in Mashen and in Large White pigs. The expression patterns of Myf5gene decreased sharply at60-day age, then keep stable lower levels afterwards. At the same stages, the relative expression quantity of Myf5gene in Mashen pig was always greater than that in Large White pig at all detecetd stages apart from that at60-day old.The expression patterns of Myf5at the levels of mRNA and protein were associated with the genetic background and the developmental stages of invididuals.