Shen Xiaopeng, Xu Feng, Wu Shen, Li Meng, Zhang Jingyi, Liang Rui, Liu Yu
School of Life Sciences, Anhui Normal University, Wuhu, China.
The Key Laboratory of Biomedicine in Gene Diseases and Health of Anhui Higher Education Institutes, Anhui Normal University, Wuhu, China.
Bio Protoc. 2019 Mar 5;9(5):e3180. doi: 10.21769/BioProtoc.3180.
In many fields of biology, especially in the field of developmental biology, reporter staining is an approach used to monitor gene expression patterns. In the reporter system, the gene is inserted in the endogenous location of the target gene via gene knock-in or by constructing a transgenic cassette in which is placed downstream of the promoter of the target gene being examined. Currently, the most common staining methods used are X-gal/FeCN staining and S-gal/TNBT staining. A serious limitation of both of these methods is that they are not effective when the gene is expressed at a low level. In an attempt to remedy this problem, we have established a new staining protocol which combines both methods. When compared to them, the method described here is better for visualizing lowly expressed genes and it has low background with high sensitivity.
在生物学的许多领域，尤其是发育生物学领域，报告基因染色是一种用于监测基因表达模式的方法。在报告基因系统中，通过基因敲入或将其构建在转基因盒中，将报告基因插入到靶基因的内源性位置，该转基因盒放置在所检测的靶基因启动子的下游。目前，最常用的染色方法是X-gal/亚铁氰化钾染色和S-gal/TNBT染色。这两种方法的一个严重局限性是，当报告基因低水平表达时它们无效。为了解决这个问题，我们建立了一种新的染色方案，该方案结合了这两种方法。与它们相比，这里描述的方法在可视化低表达基因方面更好，并且具有低背景和高灵敏度。
