【摘 要】
：
随着越来越多具有功能lncRNA被发现,IncRNA新的作用机制也逐步被揭示出来.然而,大多数lncRNA对研究中通常检测的细胞生物学效应的影响不明显,使lncRNA功能研究受到了制约.我们希望通过检测表达受到lncRNA影响的基因,进而推断出lncRNA相关功能.为此,我们将多重PCR的方法应用到了lncRNA功能研究中.多重PCR是在同一PCR反应体系里可以同时扩增多个基因片段的PCR反应.目
【机 构】
：
军事医学科学院放射与辐射医学研究所 军事医学科学院放射与辐射医学研究所;吉林大学生命科学学院
【出 处】
：
中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议
论文部分内容阅读
随着越来越多具有功能lncRNA被发现,IncRNA新的作用机制也逐步被揭示出来.然而,大多数lncRNA对研究中通常检测的细胞生物学效应的影响不明显,使lncRNA功能研究受到了制约.我们希望通过检测表达受到lncRNA影响的基因,进而推断出lncRNA相关功能.为此,我们将多重PCR的方法应用到了lncRNA功能研究中.多重PCR是在同一PCR反应体系里可以同时扩增多个基因片段的PCR反应.目前多重PCR应用于对多种病原微生物的同时检测、遗传病及癌基因的分型鉴定等,具有高效、系统、经济、简单等特点.我们选取了针对细胞DNA损伤、细胞周期凋亡、细胞信号转导及多梳蛋白基因等与癌症发生发展相关的124个重要基因,设计了35组多重PCR引物,通过优化dNTP、镁离子和DNA聚合酶的浓度及配比,使多重PCR引物在HepG2和HeLa等细胞系相关基因检测中具有良的特异性.应用上述多重PCR体系,我们对过表达及干涉特定lncRNA的细胞系进行了基因表达分析,获得了受候选lncRNA表达影响的不同生物学过程中的重要基因.利用多重PCR检测方法不仅有助于发现lncRNA分子的生物学功能,也便于揭示lncRNA在不同生物学通路中的作用.
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