中文摘要：

脂肪特异蛋白27（FSP27）是一种新发现的定位在脂滴的蛋白质,在调控脂肪细胞内脂类贮存等方面起着重要作用。本试验用含有猪FSP27基因的慢病毒,感染鼠3T3-L1前脂肪细胞,建立猪FSP27基因过表达的前脂肪细胞系,为进一步研究猪FSP27基因在脂肪细胞中调控脂类代谢提供实验素材。根据猪的FSP27基因序列设计目的基因引物序列,并分别添加酶切位点Nhe I和Age I及保护碱基,克隆到p MD18-T载体做Nhe I和Age I双酶切,将酶切产物回收并将双酶切后的目的片段与GV341载体连接,转入293T细胞,包装成慢病毒,收集、浓缩、纯化病毒上清并用来感染3T3-L1前脂肪细胞,筛选稳定系,并检测感染效率和脂肪细胞表型变化。结果显示,猪FSP27基因过表达3T3-L1脂肪细胞中FSP27基因的相对表达量显著高于对照组（P〈0.01）,在过表达脂肪细胞培养第8天,脂肪细胞形成脂滴数量明显多于对照组（P〈0.05）,表明猪FSP27基因过表达3T3-L1脂肪细胞系构建成功。

英文摘要：

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 de- signed 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 overex- pression 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 cyte cell line provides a useful tool for the study of function FSP27 gene in vivo. preadipocyte cell line provides a useful tool for the study of function FSP27 gene in vivo.