【摘要】 目的　获取人HLF细胞内稳定高表达人DNA聚合酶beta(pol β) ,并对pol β稳定高表达细胞的细胞学特征进行分析。方法　使用构建的人pol β全长cDNA的绿色荧光蛋白GFP表达载体pEGFP Cl β ,转染人HLF细胞 ,经G418筛选 ,获得单克隆的pol β稳定高表达的细胞HLFβ ,并对HLFβ细胞的细胞增殖、细胞周期、自发突变率、恶性增殖能力等特征进行分析。结果　Western印迹显示 ,HLFβ细胞的pol β蛋白表达稳定 ,较空载体转染对照细胞高 60 %以上 ,且在不同代间表达稳定。HLFβ细胞增殖能力较对照细胞增强。HLFβ细胞自发突变率略有增加 ,但与对照细胞的差异无显著性 ,细胞周期分布及细胞恶性程度亦无明显改变。结论　该研究成功获得人pol β稳定高表达细胞HLFβ ,在正常生理状况下 ,pol β的高表达对细胞周期、自发突变和恶性增殖能力未见明显影响。更多还原

【Abstract】 Objective To stably over-express human DNA polymerase beta gene(pol-β) in human HLF cells,and investigate the cellular characteristics of HLF β cells stably overxpressing human pol-β.Methods Human pol-β entire cDNA expression vector was constructed and transfected into human HLF,then screened by G418.The single cell clone with strongest fluorescence was chosen eventually and was named HLFβ.Stable overexpression of pol-β in HLFβ was confirmed by Western blot,and the biological characteristics of HLF β were analyzed from cell cycle,spontaneous mutation and cell soft agar growth.Results Expression level of pol-β in HLFβ was increased significantly over 60% as comparison with the controls,the growth speed of HLFβ appeared to be faster than that of controls.No significant change was found between HLF β and the controls in spontaneous mutation frequencies,as well as cell cycle distribution and soft agar growths.Conclusion HLFβ with overexpressing pol-β shows to be a appropriate cell line for the further study of pol-β function.更多还原