CLONING OF BASIC FIBROBLAST GROWTH FACTOR FROM CHINESE SMALL TAIL HAN SHEEP AND ITS EFFECTS ON PROLIFERATION OF MURINE C2C12 MYOBLASTS

To further understand characteristics and functions of basic fibroblast growth factor (bFGF, FGF-2) as well as its effects on skeletal muscle, a coding sequence of 18kDa-bFGF gene was cloned from Chinese Small Tail Han sheep, an excellent indigenous animal breed in China. The DNA sequence and deduced protein sequence were analyzed and aligned with the counterparts of other published animals. The results indicated that most hallmarks of the newly cloned bFGF gene sequence were found to be similar to those of published species, which implied further that bFGF genes have been conservative in evolution. The complete coding sequence for the gene was inserted into pET-28a plasmid and expressed predicted fusion protein in E. coli. The product was soluble and possessed of biological activity, it is sure to be a good source for further investigation. In addition, effects of the products on proliferation of murine C2C12 myoblasts were investigated. The findings demonstrated that the ovine recombinant bFGF accelerated in vitro the proliferation of murine C2C12 myoblasts, and MEK/ERK-MAPK signaling pathway participated in the regulative mechanism.