摘要
以绿色荧光蛋白(GFP)基因(gfp)为报告基因,构建小鼠脂联素(mADPN)基因(mAd)与gfp的融合基因mAd/gfp表达载体pCI-neo-apoEHCR-hAATp-mAd-gfp,脂质体法转染体外培养的COS-7细胞,荧光显微镜观察GFP在细胞中的表达可间接反映mADPN的表达,并通过RT-PCR在核酸水平进一步确证mAd的表达。荧光显微镜观察及RT-PCR结果均证明mADPN在COS-7细胞中获得了高效表达,表明mADPN重组表达载体pCI-neo-apoEHCR-hAATp-mAd可以在真核细胞COS-7中高效表达mADPN,为进一步探讨mAd在小鼠体内的表达提供了可行性依据。
To verify the expression of the mouse adiponeetin（mADPN） gene（mAd） recombinant expression vector pCl-neo-apoEHCR-hAATp-mAd in mammalian cells. As a reporter, the gree,a fluorescent protein（GFP） gene（gfp） was fused to the downstream of mAd, and the fusion gene mAd/gfp was cloned into the expressive vector pCl-neo-apoEHCR-hAATp to form the recombinant vector pCl-neo-apoEHCR-hAATp-mAd-gfp, which was transferred into COS-7 cells by liposome. The expression of GFP in COS-7 cells was observed by fluorescent microscopy, and the mRNA expression of mAd was detected by RT-PCR. Results showed that the recombinant mADPN was efficiently and stably expressed in COS-7 by vector pCl-neo-apoEHCR-hAATp-,nAd-gfp. The recombinant vector pCl-neo-apoEHCR-hAATp-mAd can express mADPN in mammalian cells, and it＇s proper to be used in mouse liver.
出处
《生物技术通讯》
CAS
2005年第5期492-495,共4页
Letters in Biotechnology
关键词
脂联素
绿色荧七蛋白
融合基因
真核表达
adiponectin
green fluorescent protein
fusion gene
eukaryotic expression
