Separation of Specific Cells Using a Laser Micro-dissection System and Detection of mRNA in Developing Rice Endosperm Tissue
Separation of Specific Cells Using a Laser Micro-dissection System and Detection of mRNA in Developing Rice Endosperm Tissue
复制标题
使用激光显微切割系统分离特定细胞并检测发育中的水稻胚乳组织中的 mRNA
DOI：
10.5511/plantbiotechnology.19.129
复制
发表时间：
2002
影响因子：
1.6
通讯作者：
Kunisuke Tanaka
中科院分区：
文献类型：
--
作者：
A. Kurita;R. Ohyama;S. Morita;T. Masumura;Kunisuke Tanaka
在靶细胞中克隆表达基因对于了解特异性基因表达的分子机制具有重要意义。激光显微切割技术允许从形态上不同的细胞中精确分离单个靶细胞。我们在这里描述了一种从植物组织的小目标区域分离单个细胞或由几个细胞组成的细胞簇的极好方法。在水稻胚乳组织中，糊粉层和淀粉质胚乳组织由单个胚乳母细胞分化而来。我们成功地从胚乳和糊粉层组织（开花后7天）解剖出特定的细胞区域。这些信息将有助于研究胚乳组织的细胞分化和发育机制。肌动蛋白和谷蛋白mRNA的RT-PCR分析证实了切割细胞的纯度。
Cloning of the expressing gene in the target cell is important for understanding the molecular mechanism of specific gene expression. Laser micro-dissection techniques allow the precise separation of a single target cell from morphologically distinct cells. We describe here an excellent method for separating a single cell or a cell cluster consisting of a few cells from a small target area of plant tissues. In rice endosperm tissues, the aleurone layer and starchy endosperm tissue are differentiated from a single endosperm mother cell. We succeeded in dissecting a specific cell area from endosperm and aleurone tissue (7 days after flowering). This information will be useful for studying cell differentiation and developing mechanisms of endosperm tissue. RT-PCR analysis for mRNAs for actin and glutelin confirmed the purity of the dissected cells.