人FUT3基因真核表达载体的构建与表达.docVIP

ISSN 1009-3079 CN 14-1260/R 世界华人消化杂志 2009年11月8日;17(31): 3210-3213 基础研究 BASIC RESEARCH 人FUT3基因真核表达载体的构建与表达岳丽玲, 樊 丽, 刘吉成 岳丽玲, 樊丽, 刘吉成, 齐齐哈尔医学院医药科学研究所 黑龙江省齐齐哈尔市 161006岳丽玲, 黑龙江中医药大学 黑龙江省哈尔滨市 150040岳丽玲, 在读博士, 副教授, 主要从事多糖抗肿瘤的研究.国家自然科学基金资助项目, No.黑龙江省青年科学资金资助项目, No. QC08C28作者贡献分布: 本课题由岳丽玲与樊丽设计; 研究过程由岳丽玲与樊丽操作, 刘吉成指导完成; 本论文写作由岳丽玲完成.通讯作者: 刘吉成, 教授, 博士生导师, 161006, 黑龙江省齐齐哈尔市, 齐齐哈尔医学院医药科学研究所. qyybliu@126.com电话: 0452-2663103收稿日期: 2009-09-02 修回日期: 2009-11-02接受日期: 2009-11-02 在线出版日期: 2009-11-08Construction of human FUT3 eukaryotic expression vector and its expression in human breast adenocarcinoma cells Li-Ling Yue, Li Fan, Ji-Cheng LiuLi-Ling Yue, Li Fan, Ji-Cheng Liu, Medicine and Drug Research Institute, Qiqihar Medical University, Qiqihar 161006, Heilongjiang Province, ChinaLi-Ling Yue, Heilongjiang University of Chinese Medicine, Harbin 150040, Heilongjiang Province, ChinaSupported by: National Natural Science Foundation of China, No. and the Youth Science Foundation of Heilongjiang Province, No. QC08C28Correspondence to: Professor Ji-Cheng Liu, Medicine and Drug Research Institute, Qiqihar Medical University, Qiqihar 161006, Heilongjiang Province, China. qyybliu@126.comReceived: 2009-09-02 Revised: 2009-11-02Accepted: 2009-11-02 Published online: 2009-11-08 AbstractAIM: To construct the human FUT3 (α 1, 3-fucosyltransferase) eukaryotic expression vector and analyze its expression in human breast adenocarcinoma MDA-MB-231 cells.METHODS: The full-length FUT3 cDNA was obtained by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into pMD18-T simple vector for sequence analysis. Then the FUT3 gene was subcloned into pEGFP-C1 plasmid. The resulting recombinant vector pEGFP-C1-FUT3 was identified by digestion with restriction endonucleases and transfected into MDA-MB-231 cells. A stably transfected cell line was established using G418 selection. The expression of FUT3 was observed under a fluorescence microscope and e