IGF2真核表达载体的构建及绵羊骨骼肌细胞转基因研究

Construction of the eukaryotic expression vector of IGF2 gene and transgenic research in sheep skeletal muscle cells

为了研究胰岛素样生长因子2(IGF2)对肌肉细胞生长发育的调控作用,试验对绵羊骨骼肌卫星细胞进行了分离培养,构建真核表达载体pEGFP-N1-IGF2,并转染绵羊骨骼肌细胞,然后进行半定量RT-PCR分析。结果表明:IGF2能够促进肌肉细胞的生长发育。说明建立稳定转染细胞系,进一步通过核移植可以制备IGF2转基因动物。

To study the role of insulin -like growth factor Ⅱ （IGF2） gene in the regulation of growth and development in muscle cells, the sheep skeletal muscle satellite cells were isolated and cultured. The eukaryotic expression vector pEGFP - N1 - IGF2 was constructed, and used to transfect sheep skeletal muscle cells. The semi -quantitative RT- PCR analysis was carried out. The result showed that IGF2 could promote growth and development in the muscle cells. The results illustrate that IGF2 transgenic sheep can be made using nuclear transfer technique through the establishment of stably - transfected cell lines.