| 人肿瘤MUC1/Y基因真核表达载体的构建及在BIU-87细胞中的表达 |
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| 引用本文： | 罗刚,周四维,鲁雄兵,宋晓东,孙凯,曹正国,叶章群.人肿瘤MUC1/Y基因真核表达载体的构建及在BIU-87细胞中的表达[J].武汉大学学报(医学版),2005,26(2):186-189,F002. |
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| 作者姓名： | 罗刚 周四维 鲁雄兵 宋晓东 孙凯 曹正国 叶章群 |
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| 作者单位： | 华中科技大学同济医学院附属同济医院泌尿外科,武汉,430030;华中科技大学同济医学院附属同济医院泌尿外科,武汉,430030;华中科技大学同济医学院附属同济医院泌尿外科,武汉,430030;华中科技大学同济医学院附属同济医院泌尿外科,武汉,430030;华中科技大学同济医学院附属同济医院泌尿外科,武汉,430030;华中科技大学同济医学院附属同济医院泌尿外科,武汉,430030;华中科技大学同济医学院附属同济医院泌尿外科,武汉,430030 |
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| 基金项目： | 国家自然科学基金资助项目(编号:30271300) |
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| 摘 要： | 目的:克隆人肿瘤MUC1/Y全长基因及构建真核表达载体 pEGFP MUC1/Y并观察 pEGFP MUC1/Y在 BIU 87细胞中表达以用于进一步生物学功能及肿瘤生物学治疗的研究。方法: 取新鲜膀胱肿瘤组织提取总RNA,采用随机引物进行反转录,将特异引物扩增的MUC1/Y全长 PCR产物克隆至真核表达载体 pEGFP C1,测序鉴定后命名为 pEGFP MUC1/Y,用脂质体转染膀胱肿瘤细胞BIU 87,以 Western Blot检测其表达情况。结果:酶切鉴定和序列分析证实构建质粒含人MUC1/Y全长cDNA编码序列, Western Blot检测和转染实验,表明构建的 pEGFP MUC1/Y基因在BIU 87真核细胞中表达,其绿色荧光蛋白瞬时表达率为 20%。结论:人肿瘤 MUC1/Y全长cDNA基因克隆及其真核表达载体 pEGFP MUC1/Y 构建成功,可用于肿瘤生物学治疗的研究。
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| 关 键 词： | MUC1/Y 克隆 绿色荧光蛋白 真核表达 |
| 文章编号： | 1671-8852(2005)02-0186-04 |
Construction of Eukaryotic Expression Vector EGFP-MUC1/Y and Its Expression in Bladder Carcinoma Cell Line BIU-87 |
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| Luo Gang,Zhou Siwei,Lu Xiongbing,et al Dept. of Urology,Tongji Hosipital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan ,China.Construction of Eukaryotic Expression Vector EGFP-MUC1/Y and Its Expression in Bladder Carcinoma Cell Line BIU-87[J].Medical Journal of Wuhan University,2005,26(2):186-189,F002. |
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| Authors: | Luo Gang Zhou Siwei Lu Xiongbing Dept of Urology Tongji Hosipital Tongji Medical College Huazhong University of Science and Technology Wuhan China |
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| Institution: | Luo Gang,Zhou Siwei,Lu Xiongbing,et al Dept. of Urology,Tongji Hosipital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China |
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| Abstract: | Objective: To clone human full length MUC1/Y cDNA and construct an eukaryotic expression vector EGFP-MUC1/Y, then transfect it into bladder carcinoma cell line BIU-87 and detect its expression with Western blot and green fluorescent protein. Methods: The total RNA of human bladder carcinoma was extracted and amplified by RT-PCR primed by random and specific primers respectively. The amplified products were cloned and linked in green fluorescent protein vector EGFP-C1. The recombinant EGFP vector was transfected into human bladder tumor cell BIU-87 by Lipofectamine.Results: The cDNA sequencing showed that the recombinant plasmid contained the coding region of human full length MUC1/Y gene and could be expressed in 20% of BIU-87 cells. Conclusion: The human full-length MUC1/Y gene was cloned and expressed in human bladder carcinoma cell line BIU-87, and the results indicates it has potential roles in tumor gene therapy. |
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| Keywords: | MUC1/Y Clone Green Fluorescent Protein Eukaryotic Expression |
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