| pEGFP-N1-UCH-L1重组质粒的构建 |
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| 引用本文： | 卢豪,;庞伟,;徐婷,;杨红澎,;奚用勇,;黄承钰,;蒋与刚. pEGFP-N1-UCH-L1重组质粒的构建[J]. 西南国防医药, 2014, 0(8): 813-815 |
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| 作者姓名： | 卢豪, 庞伟, 徐婷, 杨红澎, 奚用勇, 黄承钰, 蒋与刚 |
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| 作者单位： | [1]成都军区疾病预防控制中心,成都610021; [2]军事医学科学院卫生学环境医学研究所营养研究室;,成都610021; [3]成都市第二人民医院;,成都610021; [4]四川大学华西公共卫生学院营养与食品卫生教研室,成都610021; |
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| 基金项目： | 国家自然科学基金（30872098,30901185）;天津市自然科学基金（05YFJMJC05500）;全军医学科技青年培育项目（13QNP069） |
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| 摘 要： | 目的 构建pEGFP-N1-UCH-L1重组质粒,并进行鉴定。方法提取Wistar乳鼠大脑组织总RNA,逆转录合成cDNA,经PCR扩增UCH-L1基因,定向克隆至载体pEGFP-N1,构建重组质粒,经测序进行鉴定。结果 UCH-L1基因重组质粒经测序鉴定证明构建正确。结论成功构建了UCH-L1基因重组质粒,可用于神经细胞转染,进行后续研究。
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| 关 键 词： | 泛素羧基末端水解酶L1 pEGFP-N1载体 重组质粒 |
Construction of recombinant plasmid pEGFP-N1-UCH-L1 |
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| Affiliation: | Lu Hao , Pang Wei, Xu Ting, Yang Hongpeng, Xi Yongyong, Huang Chengyu, Jiang Yugang( 1. Centre for Disease Control and Prevention, Chengdu Military Command, Chengdu, Sichuan, 610021, China; 2. Department of Nutrition, Institute of Health ＆ Environmental Medicine, Academy of Military Medical Sciences, Tianjin, 300050, China; 3. Second People ＂s Hospital of Chengdu, Sichuan,610000, China; 4. Department of Nutrition and Food Hygiene, West China School of Public Health, Sichuan University, Chengdu, Sichuan,610041, China) |
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| Abstract: | Objective To construct and identify the eukaryotic expression vector for UCH-L1 gene. Methods Total RNA was extracted from newborn Wistar rats and reversely transcribed to cDNA,with which UCH-L1 gene was amplified by RT-PCR and cloned into the eukaryotic expression vector pEGFP-N1; the constructed recombinant plasmid was identified by restriction analysis and sequencing. Results The results of restriction analysis and sequencing proved that UCH-L1 gene is successfully inserted into plasmid pEGFP-N1. Conclusions The eukaryotic expression vector for UCH-L1 is successfully constructed,and can be used to transfect neurons in further study. |
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| Keywords: | UCH-L1 pEGFP-N1 vector recombinant plasmid |
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