Pluskota Wioletta E, Bradford Kent J, Nonogaki Hiro
Department of Plant Physiology and Biotechnology, University of Warmia and Mazury, Olsztyn, Poland.
Methods Mol Biol. 2011;773:329-39. doi: 10.1007/978-1-61779-231-1_19.
A number of genes and proteins are expressed in a tissue- or cell layer-specific manner. Spatial patterns of gene expression are critical to understanding gene function. Tissue printing provides a simple and rapid method to analyze localization of mRNA and protein at the tissue and cellular levels. This is especially convenient for gene expression analysis in hard tissues, such as seeds that are often difficult to section. Seed RNA or protein can be transferred onto a suitable membrane by printing the cut surface of a bisected seed. This method has been used successfully to determine mRNA and protein localization in seed research. The resolution of printed seed images and RNA and protein signals in tissue printing is sufficient to identify embryo- or endosperm-specific expression of various genes and proteins. In some cases, these studies have contributed to elucidating the spatial characteristics of hydrolytic enzymes putatively involved in the completion of germination and/or early postgerminative growth. By the same principle, tissue-printing methods could also be valuable for elucidating the spatial characteristics of genes/proteins that control the inception, maintenance, and termination of seed dormancy.
许多基因和蛋白质以组织或细胞层特异性的方式表达。基因表达的空间模式对于理解基因功能至关重要。组织印迹提供了一种简单快速的方法来分析mRNA和蛋白质在组织和细胞水平上的定位。这对于硬组织中的基因表达分析特别方便，例如种子，种子通常很难切片。通过将对半切开的种子的切割面进行印迹，可以将种子RNA或蛋白质转移到合适的膜上。该方法已成功用于种子研究中确定mRNA和蛋白质的定位。组织印迹中打印种子图像以及RNA和蛋白质信号的分辨率足以识别各种基因和蛋白质在胚或胚乳中的特异性表达。在某些情况下，这些研究有助于阐明可能参与发芽完成和/或发芽后早期生长的水解酶的空间特征。基于同样的原理，组织印迹方法对于阐明控制种子休眠起始、维持和终止的基因/蛋白质的空间特征也可能具有重要价值。
