Establishment and Verification of Porcine FSP27 Gene Overexpression Preadipocyte

Fat-specific protein 27 (Fsp27),localizing in the surface of lipid droplets(LDs),plays an important role in controlling lipid store in adipocytes. In this study, the containing porcine FSP27 gene of lentiviral vector was used to infect mouse 3T3-L1 preadipocytes which provide experimental material for further study of FSP27 gene regulation lipid metabolism in adipocytes. Primer sequences were designed based on FSP27 gene and their restriction sites were added with NheI and AgeI, and then inserted into pMD18-T vector. pMD18-T-FSP27 was connected with GV341 vector that co-transfected into 293T cells with the HA-mFSP27 expression vector to test its expression efficiency. Then, the FSP27 overexpression vector was transferred to a lentiviral vector. Lentivirus were generated and used to infect 3T3-L1 cells. The results showed that the relative expression level of FSP27 gene in FSP27 gene overexpression preadipocyte cell was significantly higher than control group (P 0.01). And, the number of LDs in FSP27 gene overexpression adipocytes cultured 8 days was significantly higher than control group(P 0.05). It is suggested that the successful construction of porcine FSP27 gene overexpression preadipocyte cell line provides a useful tool for the study of function FSP27 gene in vivo.