摘要
利用分子克隆技术,采用RT-PCR扩增follistatin related-gene全长cDNA序列,克隆入真核表达载体pEGFP-N1后测序鉴定,并通过脂质体瞬时转染小鼠成肌细胞,采用RT-PCR和SDS-PAGE方法鉴定FLRG蛋白表达情况。结果表明,成功构建的真核表达载体pEGFP-N1-FLRG能成功转染小鼠成肌细胞,RT-PCR和SDS-PAGE电泳证实pEGFP-N1-FLRG在成肌细胞中表达出FLRG蛋白,为深入研究follistatin related-gene奠定了基础。
The full length cDNA of follistatin related-gene was amplified through reverse transfer polymerase chain reaction（RT- PCR）and subcloned into eukaryotic expression vector pEGFP-N1. The recombinant plasmid pEGFP-N1-FLRG was transfected into mouse myoblast cells with liposome. The expression of mRNA and protein encoded by this gene was detected respectively with RT-PCR and SDS-PAGE. The result showed that follistatin related-gene was cloned into pEGFP-N1 successfully, and its expression at mRNA and protein level was detected with RT-PCR and SDS-PAGE. The experiment established the basis for further study on the function of follistatin related-gene.
出处
《沈阳农业大学学报》
CAS
CSCD
北大核心
2008年第2期243-246,共4页
Journal of Shenyang Agricultural University
关键词
FLRG
转染
真核表达
FLRG
transfection
eukaryotic expression
