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作 者:谭琛[1] 董利[1] 李江[1] 曹利[1] 彭聪[1] 李小玲[1] 李桂源[1]
机构地区:[1]中南大学湘雅医学院肿瘤研究所,中国湖南长沙410078
出 处:《生命科学研究》2002年第1期74-78,共5页Life Science Research
基 金:国家自然科学基金资助项目 ( 3980 0 14 2 ;30 10 0 10 5 )
摘 要:为了研究NAG7基因编码产物的特性和在活细胞内定位与表达 ,首先利用生物信息学分析NAG7编码蛋白的一般性质并预测其定位 ,再通过构建增强型绿色荧光蛋白 (Enhancegreenfluorescentprotein ,EGFP)与NAG7融合基因的真核表达载体pEGFP C2 NAG7,通过脂质体介导分别转染非洲绿猴肾细胞系COS7和人鼻咽癌细胞系HNE1 ,瞬时表达后荧光显微镜观察NAG7基因编码蛋白的活细胞内定位及表达 .研究结果表明NAG7的编码产物可在COS7细胞中高表达并定位于细胞浆 ,而在HNE1细胞中虽也定位于胞浆 ,但仅有极少数细胞存在表达 。To analyse the normal expression and location of NAG7 coding protein and to understand the relationship between the expression pattern of the protein and cell carcinogenesis, bioinformatics was used to assay the NAG7 protein character to provide an available clue for subsequent research. The NAG7 codon frame cDNA was amplified by PCR, and subcloned into enhance green fluorescence gene (EGFP). The recombinant plasmid was transfected into HNE1 and COS-7 cells. The expression of NAG7 coding protein was observed by fluorescence microscopy. The results demonstrated that the expressed GFP-NAG7 fusion protein generated striking green fluorescence in the cytoplasm in HNE1 and COS-7 cells. Green fluorescence was generated in most of COS-7 cells but in a few HNE1 cells, which suggested that the expressional difference in HNE1 might involve in the carcinogenesis of NPC.
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