含人过氧化物酶体增殖物激活受体δ基因高效真核表达载体的构建及其意义

章涛;杨贵忠;袁野 ;万敬员;杨俊卿 ;蒋建新 ;周岐新 

解剖学报 ›› 2007, Vol. 38 ›› Issue (2) : 173-177.

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解剖学报 ›› 2007, Vol. 38 ›› Issue (2) : 173-177.
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含人过氧化物酶体增殖物激活受体δ基因高效真核表达载体的构建及其意义

  • 章涛 1; 杨贵忠 1; 袁野 1 ;万敬员1 ;杨俊卿 1 ;蒋建新 2 ;周岐新 1* BR>
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THE CONSTRUCTION AND SIGNIFICANCE OF THE HIGHLY EFFICIENT EUKARYOTIC EXPRESSION VECTOR CARRYING HUMAN PPARδ GENE

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Abstract

Objective To construct a highly efficient eukaryotic expression vector carrying human peroxisome proliferatoractivated receptor δ(hPPARδ) gene in order to provide an ideal molecular platform for screening natural ligands and functional study of hPPARδ. Methods hPPARδ gene, cloned from total RNA of HepG2 cells by RT-PCR, was ligated with pIRES2EGFP plasmid which was excised by BamHI and SalI double endonucleases. The recombinant plasmid was transfected into 293 cells. Realtime quantitative PCR and immunocytochemistry assays were used to analyze the expression levels of hPPARδ in the transfected 293 cells. Results hPPARδ gene sequence contained in the recombinant plasmid phPPARδIRES2EGFP was verified correct by enzyme digestion as well as sequence analysis. After being transfected into 293 cells, high efficient expression of hPPARδ gene contained in phPPARδIRES2EGFP plasmid was found to display a highly efficient expression detected both at mRNA and protein levels by r

关键词

过氧化物酶体增殖物激活受体δ / 基因克隆 / 真核表达 / 逆转录-聚合酶链反应

Key words

Peroxisome proliferatoractivated receptor δ / Gene cloning / Eukaryotic expression / RT-PCR

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章涛;杨贵忠;袁野 ;万敬员;杨俊卿 ;蒋建新 ;周岐新 . 含人过氧化物酶体增殖物激活受体δ基因高效真核表达载体的构建及其意义[J]. 解剖学报. 2007, 38(2): 173-177
THE CONSTRUCTION AND SIGNIFICANCE OF THE HIGHLY EFFICIENT EUKARYOTIC EXPRESSION VECTOR CARRYING HUMAN PPARδ GENE[J]. Acta Anatomica Sinica. 2007, 38(2): 173-177
中图分类号: R34    R966   

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