Quantitative measurement of mRNA at different loci within an individual living cell.
Quantitative measurement of mRNA at different loci within an individual living cell.
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DOI：
10.2142/biophys.43.s161_3
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发表时间：
2004-08
期刊：
影响因子：
2.2
通讯作者：
H. Uehara;T. Osada;A. Ikai
中科院分区：
文献类型：
--
作者：
H. Uehara;T. Osada;A. Ikai
细胞蛋白质和mRNA的不对称定位对于细胞功能如分裂、分化和发育是重要的。特定mRNA的定位通过控制特定蛋白质的翻译位点从而将它们的位置限制在适当的细胞区域来产生细胞极性。我们之前曾报道过一种基于原子力显微镜（AFM）的新方法，可以在不杀死或破坏单个活细胞的情况下检测单个活细胞中的基因表达。将原子力显微镜探针插入活细胞中提取mRNA，并在增殖后通过RT-PCR和定量PCR进行分析。通过应用这种方法，在这项研究中，我们进行了定量测量mRNA在不同的位点在个别活细胞。
Asymmetric localizations of cellular proteins and mRNAs are important for cell functions such as division, differentiation and development. The localization of specific mRNA generates cell polarity by controlling the translation sites of specific proteins and thereby restricting their locations to appropriate cellular regions. We have previously reported a novel method based on atomic force microscopy (AFM) for examining gene expression in a single living cell without killing or destroying it. An AFM tip was inserted into a living cell to extract mRNAs, which were analyzed after multiplication by RT-PCR and quantitative PCR. By applying this method, in this study we performed quantitative measurement of mRNA at different loci within individual living cells.