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。
Objective To construct hPARPl-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 hPARPl-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 hPARPl-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 hPARPl-deficient strain offered an effective cell strain for studying the function of hPARP1 gene.
出处
《卫生研究》
CAS
CSCD
北大核心
2005年第6期650-652,共3页
Journal of Hygiene Research
基金
"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)
