中国生物工程杂志

To generate recombinant adenovirus expression vector of human Sema4C gene and observe its expression in mouse myoblasts cell line C2C12 for ensuring easy access to investigate the role of Sema4C gene during myogenesis. The recombinant plasmid was packaged and amplified after being transfected in HEK293 cells through Lipofectamine. After infecting C2C12 myoblasts with recombinant adenovirus vector, the adenoviral infection efficiency was determined by confocal microscope which showed that the expression of green fluorescence could be detected at 12h and then reached peak at 24h after recombinant adenovirus infection. The infection efficiency was almost 100% confirmed by FACS examination. Detection of WB indicated that the expression of Sema4C in C2C12 of recombinant adenoviral infection group was significantly higher than that of the control group (P<0.01). To further investigate the role of Sema4C gene during the proliferation and differentiation of C2C12 myoblasts, proliferation index of C2C12 cells were detected by FACS and the myogenic differentiation was also observed after adenovirus infection. Our data firstly showed that overexpression of exogenous human Sema4C gene could not only increase the percentage of G0/G1 cell cycle and decrease the proliferation index but also promote myotube formation of C2C12 cells.