Abstract:

To construct the skeletal muscle-specific FSⅠ-Ⅰ eukaryotic expression vector and identify the integration of FSⅠ-Ⅰ gene in porcine embryonic fibroblasts,and to provide a basis for study on  the effect  of FSⅠ-Ⅰ on skeletal muscle.Methods The FS A (containing FS N and FSⅠ) and FSⅠ were amplified by RT-PCR from porcine skeletal muscle;the MCK enhancer,ACTA1 promoter and BGHpolyA were amplified by PCR from mouse genenome,human genenome and pcDNA3.1(+), respectively.The  four fragments mentioned above were subcloned into the eukaryotic expression vector pGKneotpAlox2,and then the vector was transfeced into porcine embryonic fibroblasts  using the FuGENER　HD reagent,according to the manufacturer’s protocol.The cells were selected with G418 antibiotic and identified by PCR amplification.Results The FS Ⅰ-Ⅰ skeletal muscle -specific expression vector was successfully constructed  and integrated to the genome of the porcine fibroblasts and  the clone cells integrating the FSⅠ-Ⅰ gene were obtained.Conclusion The porcine fibroblast clone  expressing FSⅠ-Ⅰ  is obtained and it should be of great value to the construction of FSⅠ-Ⅰ transgenic swine.

Key words: follistatin；porcine embryonic fibroblasts；eukaryotic expression