摘要
采用RT-PCR法对海兰鸡MyoD全长cDNA序列进行扩增,克隆、测序后,构建真核表达载体pEGFP-N1-MyoD,脂质体瞬时转染鸡胚成纤维细胞,用荧光显微镜、RT-PCR和SDS-PAGE电泳法检测MyoD蛋白表达情况。结果表明,构建的真核表达载体pEGFP-N1-MyoD能成功转染鸡胚成纤维细胞;荧光显微镜观察可见绿色荧光;RT-PCR可见目的条带;SDS-PAGE电泳证明表达的蛋白相对分子质量为33KD,该结果为研究MyoD蛋白的功能奠定了试验基础。
MyoD cDNA was amplified through reverse transfer polymerase chain reaction（RT-PCR）,and then cloned and sequenced.The eukaryotic expression vector pEGFP-N1-MyoD was constructed,and was transfected into chick embryo fibroblast with liposome.The MyoD protein encoded by this gene was detected respectively with fluorescence microscope,RT-PCR and SDS-PAGE.The results showed that eukaryotic expression vector pEGFP-N1-MyoD was transfected into chick embryo fibroblast successfully.The green fluorescence was observed under a fluorescence microscope,the objective band was detected with RT-PCR and the MyoD protein was detected with SDS-PAGE,which was about 33KD.The results provided experimental basis for the function of MyoD protein.
出处
《沈阳农业大学学报》
CAS
CSCD
北大核心
2010年第1期106-109,共4页
Journal of Shenyang Agricultural University
关键词
MYOD
转染
真核表达
MyoD
transfection
eukaryotic expression
