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| RNA干扰技术建立人PARP-1缺陷细胞株 | |
| 引用本文： | 沙焱,曾玉云,庄志雄,何云,胡大林,胡恭华,杨建平,涂晓志.RNA干扰技术建立人PARP-1缺陷细胞株[J].卫生研究,2005,34(6):650-652. |
| 作者姓名： | 沙焱 曾玉云 庄志雄 何云 胡大林 胡恭华 杨建平 涂晓志 |
| 作者单位： | 1. 中山大学公共卫生学院,广州 510089 2. 深圳沙井卫生监督所 3. 深圳市疾病预防控制中心 |
| 基金项目： | “973”国家重点基础研究发展计划基金资助项目(No.2002CB512904,2002CB512903),国家自然科学基金项目(No.30571592) |
| 摘 要： | 目的建立并鉴定聚ADP核糖聚合酶1(hPARP1)缺陷细胞株,用于研究hPARP1基因的作用机制及其缺陷与DNA损伤发生的关系。方法将用已经构建的pEGFPC1shRNA(包含两个PARP1基因及一个阴性对照)转染支气管上皮细胞(16HBE),使之在16HBE中表达,转染后命名为16HBEP1、16HBEP2及16HBEN。用蛋白免疫印迹法鉴定转染细胞中hPARP1基因的表达水平。结果pEGFPC1shRNA在真核细胞成功表达;16HBEP1和16HBEP2的蛋白表达水平分别下降了84.3%及63.7%。结论hPARP1缺陷细胞株的成功建立和鉴定为hPARP1基因功能研究提供了一种有效手段。 |
| 关 键 词： | RNA干扰 短发夹-RNA hPARP1 绿色荧光蛋白载体 真核细胞转染 |
| 文章编号： | 1000-8020（2005）06-0650-03 |
| 收稿时间： | 2005-02-25 |
| 修稿时间： | 2005年2月25日 |
Construction of hPARP1-deficient cell strain by RNA interference |
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| Sha Yan,Zeng Yu-yun, Zhuang Zhi-xiong, He Yun,et al..Construction of hPARP1-deficient cell strain by RNA interference[J].Journal of Hygiene Research,2005,34(6):650-652. | |
| Authors: | Sha Yan Zeng Yu-yun Zhuang Zhi-xiong He Yun |
| Institution: | Faeuhy of Preventive Medicine, School of Public Health, Sun Yet-san University, Guangzhou 510089, China |
| Abstract: | OBJECTIVE: To construct hPARP1-deficient cell strain which can be used in studying the functions and action mechanisms of hPARP1 gene. METHODS: 16-HBEs were transfected with two kinds of eukaryotic expression plasmids of hPARP1 gene short hairpin RNA to construct hPARP1-deficient cells (named as "16-HBEP1 and 16-HBEP2") and a negative control plasmids (named as 16-HBEN). The protein expression levels of hPARP1 gene in 16-HBE, 16-HBEP1 16-HBEP2 and 16-HBEN were detected by the western blotting to estimate the effects of RNA interference. RESULTS: The construction of hPARP1-deficient cells train was successful and the protein expression level of hPARP1 gene in the 16-HBEP1 and 16-HBEP2 were decreased 84.3% and 63.7% respectively as compared with that in 16-HBE. CONCLUSION: The successes in constructing hPARP1-deficient strain offered an effective cell strain for studying the function of hPARP1 gene. |
| Keywords: | RNA interference hPARP1 short hairpin RNA green fluorescent protein vector eukaryotic transfection |
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