关键词: hNRDRA2, 核定位信号, 亚细胞定位, 绿色荧光蛋白, 重组表达

Abstract: BACKGROUND ＆ AIM: To analyze the exogenous expression and localization of hNRDRA2 in eukaryocyte and demonstrate the function of the predicted nuclear localization signal (NLS). MATERIALS AND METHODS: hNRDRA2 cDNA and NLS sequence were cloned into pEGFP-C1 to construct mammalian expression vectors pEGFP-C1-A2 and pEGFP-C1-NLS fused with green fluorescent protein (GFP). Then these constructed vectors were transiently transfected into SK-N-SH, KP-N-NS and COS-7 cells. The transfected cells were examined under fluorescent microscope. RESULTS: The recombinant plasmids were identified by sequencing. In the transfected cells, recombinant protein GFP-A2 was distributed throughout the cell, while GFP-NLS was detected mainly in the nucleus. CONCLUSION: The predicted NLS could import the fusion protein GFP into nucleus, but exogenous expressed hNRDRA2 was not localized in the nucleus.

Key words: hNRDRA2, NLS, subcellular localization, green fluorescent protein, recombinant expression