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| 肝细胞癌高表达基因TPT1真核报告表达载体的构建及鉴定 | |
| 引用本文： | 高天慧,段芳龄,马军,周云,孙嫣,孙艳,王晓,薛乐勋.肝细胞癌高表达基因TPT1真核报告表达载体的构建及鉴定[J].胃肠病学和肝病学杂志,2005,14(3):268-271. |
| 作者姓名： | 高天慧 段芳龄 马军 周云 孙嫣 孙艳 王晓 薛乐勋 |
| 作者单位： | 1. 郑州大学消化疾病研究所,郑州,450003 2. 河南省人民医院肿瘤科 3. 河南省中医学院一附院 4. 郑州大学分子生物学重点实验室 |
| 基金项目： | 河南省医学科技人才创新工程项目，国家"211"工程建设项目 |
| 摘 要： | 目的构建TPT1真核报告表达载体pEGFP-N3TPT1,为进一步研究其功能奠定基础.方法通过PCR和酶切的方法获取具有黏性末端的TPT1基因的全长开放读码框,将其定向连入真核表达载体pEGFP-N3绿色荧光报告基因的氨基侧,构成pEGFP-N3TPT1的真核报告表达载体,通过酶切和测序鉴定TPT1的正确插入.结果酶切和测序均证实TPT1正确插入了真核报告表达载体pEGFP-N3TPT1中.结论成功构建了TPT1的真核报告表达载体pEGFP-N3TPT1. |
| 关 键 词： | 真核表达 报告载体 构建 |
| 文章编号： | 1006-5709(2005)03-0268-04 |
| 修稿时间： | 2005年3月8日 |
Construction and identifying of TPT1 eukaryotic expression reporter vector |
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| Gao Tianhui,DUAN Fangling,MA Jun,ZHOU Yun,SUN Yan,SUN Yan,WANG Xi ao Institube of digestive disease,Zhengzhou university,Zhenghzou ,Ch ina.Construction and identifying of TPT1 eukaryotic expression reporter vector[J].Chinese Journal of Gastroenterology and Hepatology,2005,14(3):268-271. | |
| Authors: | Gao Tianhui DUAN Fangling MA Jun ZHOU Yun SUN Yan SUN Yan WANG Xi ao Institube of digestive disease Zhengzhou university Zhenghzou Ch ina |
| Affiliation: | Gao Tianhui,DUAN Fangling,MA Jun,ZHOU Yun,SUN Yan,SUN Yan,WANG Xi ao Institube of digestive disease,Zhengzhou university,Zhenghzou 450003,Ch ina |
| Abstract: | Objective To construct the eukaryotic expression reporter vector of TPT1. Methods The specific recognize sites of EcoRI and BamHI were introduced respectively into the two ends of TPT1 ORF through PCR met hod. TPT1 ORF PCR products and the vector pEGFP-N3 were then double diges t ed by EcoRI and BamHI restriction enzyme respectively and then ligated together. Kanamycin was used to screen the positive reconstituted vectors, plasmids were extracted by miniprep and checked through PCR and restriction endonuclease diges tion,and sequenced using Sanger method. VecScreen and BLA ST were used to analyse the sequence.Results TPT1 ORF is inserted correctly (the direction,site,and the sequence of TPT1 OR F are all correct) into the pEGFP-N3.Conclusions TPT1 eukaryotic expression reporter vector pEGFP-N3TPT1 was correctly cons tructed. |
| Keywords: | Eukaryotic expression Reporter vector Const ruction |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |
