人鼻咽组织特异性表达基因NASG编码产物在细胞中的融合表达

The Fusion Expression in Cells of NASG Coding Protein,a Tissue-specific Molecule in Human NasopharyngealEpithelial Tissue

采用PCR技术从人鼻咽上皮组织cDNA文库扩增出人鼻咽组织特异性表达基因NASG基因的编码序列,用XhoI和SalI酶切NASG基因编码序列的PCR产物及pEGFP C2载体,产生匹配的粘末端.将回收的pEGFP C2载体分别与NASG基因编码区酶切片段连接,构建了其编码产物的融合蛋白表达载体pEGFP C2 NASG,通过脂质体介导分别转染非洲绿猴肾永生化细胞系COS7和鼻咽癌细胞系HNE1,瞬时表达后荧光显微镜观察NASG编码蛋白的活细胞定位,结果表明,NASG编码蛋白在COS7细胞和HNE1细胞的胞浆和胞核中均有表达.

The coding region with Xho I and Sal I restriction sites of NASG gene,a tissuespecific gene in human nasopharyngeal epithelial tissue,was amplified by PCR from human nasopharyngeal epithelial cDNA library.The PCR product and plasmid pEGFPC2 were digested by Xho I and Sal I restrict endonucleases,then ligated by T4 DNA ligase to construct recombinant expression vector.Endonuclease digesting and DNA sequencing confirmed that the coding region of NASG gene was correctly inserted into the vector.The recombinant plasmid pEGFPC2/NASG was introduced into COS7 cells and HNE1 cells by liposome transfection respectively.Observation under the fluorescent microscopy after 24 hours of transfection showed that EGFP/NASG expressed and existed both in the cytoplasm and in the nucleus.