人STRA6基因克隆及其在肝癌细胞中的表达定位
Human STRA6 Gene Cloning and Its Expression and Localization at Hepatoma Cells
DOI: 10.12677/BIPHY.2021.91003, PDF,    国家自然科学基金支持
作者: 何建*, 王金丽*, 余畅, 陈相屹, 陈 帅, 严辉, 石静#, 孙达权#：贵州医科大学基础医学院，贵州 贵阳
关键词: 人STRA6基因；基因克隆；肝癌细胞；蛋白定位；Human STRA6 Gene； Gene Cloning； Liver Cancer Cell； Protein Localization
摘要: 克隆人STRA6基因蛋白编码区，构建STRA6基因真核表达载体并表达于肝癌细胞，分析其在肝癌细胞中的定位。用TRIzol法从人肝细胞QSG-7701中抽提获得总RNA，经RT-PCR和高保真PCR获得人STRA6基因蛋白编码区，并进行测序鉴定；用基因工程技术构建真核表达载体pDsRed1-STRA6，用脂质体法转染人肝癌细胞Hep 3B；蛋白免疫印迹检测肝癌细胞中STRA6蛋白的表达情况，激光共聚焦显微镜分析STRA6在肝癌细胞中的分布。成功克隆人STRA6基因的蛋白编码区，构建真核表达载体pDsRed1-STRA6并使其表达于肝癌细胞中，发现STRA6在肝癌细胞中表达后集中于细胞表面一个区域。本研究成功克隆STRA6基因并证明其在肝癌细胞表面分布具有区域性。
Abstract: To clone human STRA6 gene and construct its eukaryotic expressional vector to express target gene and to investigate its localization in hepatocellular carcinoma cells (HCCs). Total RNA was extracted from human liver cells QSG-7701 by TRIzol method, and the coding region of human STRA6 gene, identified by DNA sequencing, was obtained by RT-PCR and high fidelity PCR. Then, the eukaryotic expressional vector pDsRed1-STRA6 was constructed by genetic engineering technology and transfected into human hepatoma cell Hep 3B by liposome method. The expression of STRA6 protein in HCC cells was detected by Western blot, and the distribution of STRA6 in HCC cells was analyzed by laser scanning confocal microscopy. The coding region of human STRA6 gene was cloned successfully, and the eukaryotic expressional vector pDsRed1-STRA6 was constructed and expressed in HCC cells. It was found that the expressed STRA6 in HCC cells concentrated in one region on the cell surface. The STRA6 gene was successfully cloned and its protein distributed regionally on the surface of hepatoma cells.
文章引用：何建, 王金丽, 余畅, 陈相屹, 陈帅, 严辉, 石静, 孙达权. 人STRA6基因克隆及其在肝癌细胞中的表达定位[J]. 生物物理学, 2021, 9(1): 22-28. https://doi.org/10.12677/BIPHY.2021.91003

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