牛畸胎瘤衍化生长因子TDGF1基因过表达对成肌细胞增殖的影响

Expression of bovine TDGF1 gene and its effects on myoblast proliferation

为明确牛畸胎瘤衍化生长因子(TDGF1)对成肌细胞(C2C12)增殖的影响,利用定向克隆和细胞转染技术获得可稳定过表达牛TDGF1基因的C2C12细胞系,Western blot(WB)和荧光定量PCR技术检测TDGF1在C2C12细胞系中的表达量,MTT法检测C2C12细胞的增殖。结果表明,成功构建了pIRES2-EGFP-TDGF1真核表达载体,牛TDGF1基因在C2C12细胞中的表达量显著上调,并成功表达出TDGF1蛋白,且过表达牛TDGF1基因可抑制C2C12细胞增殖,在转染30h时抑制作用最显著。

In order to finding out that the bovine TDGF1 affects myoblast proliferation,using the directional cloning and cell transfection technique to obtain stable expression of bovine TDGF1 gene in C2C12 cell line,Western blot（WB） and fluorescence quantitative PCR technology detected TDGF1 in the expression of C2C12 cell line,MTT method detected C2C12 cells proliferation. The result indicated that the pIRES2-EGFP-TDGF1 eukaryotic expression vector was obtained success- fully,bovine TDGF1 gene was expressed significantly in C2C12 cells,and TDGF1 protein was ex- pressed successfully,bovine TDGF1 gene could inhibit the myoblast proliferation and especially at 30 h after transfection.