摘要
用RT-PCR扩增含有XhoI/EcoRI酶切位点的Sirt1片段,克隆、测序后,构建真核荧光表达载体pEGFP-N1-Sirt1、脂质体介导转染293细胞,最后利用SDS-PAGE检测蛋白表达情况。结果表明,经脂质体介导重组体pEGFP-N1-Sirt1可成功转染293细胞;转染48h后,荧光显微镜下可见绿色荧光蛋白表达;SDS-PAGE证明表达的融合蛋白相对分子质量约为41kD。该结果为进一步研究Sirt1在哺乳动物细胞中的作用奠定了基础。
Sirt1 eDNA containing XhoI/EcoRI sites was amplified by RT-PCR, and then cloned and sequenced. Eukaryotic expression vector pEGFP-N1-Sirtl was transfected into 293 cells with lipidosome, and the expression protein was detected by SDSPAGE. The results showed that pEGFP-N1-Sirt1 could be transfected into 293 cells with lipidosome successfully. The green fluorescence protein was observed 48h after transfection. The fusion protein was detected with SDS-PAGE, which was about 41kD. The results provided experimental basis for further study on the function of Sirtl gene.
出处
《沈阳农业大学学报》
CAS
CSCD
北大核心
2008年第6期695-698,共4页
Journal of Shenyang Agricultural University
基金
辽宁省自然科学基金资助项目(20050302)
辽宁省教育厅科研基金资助项目(2007304)
