JK
Joseph C. Kuhl
  • Department of Plant Sciences, University of Idaho, Moscow
研究方向
  • Molecular biology
In situ Hybridization of Plant-parasitic Nematode Globodera pallida Juveniles to Detect Gene Expression
原位杂交检测马铃薯白胞囊线虫的基因表达
作者:Joanna Kud, Nejra Solo, Allan Caplan, Joseph C. Kuhl, Louise-Marie Dandurand and Fangming Xiao日期:09/20/2019,浏览量:4000,Q&A: 0
In this study, we describe a standard whole mount in situ hybridization method which is used to determine the spatial-temporal expression pattern of genes from Globodera spp. Unlike more invasive radioactive labeling approaches, this technique is based on a safe, highly specific enzyme-linked immunoassay where a Digoxigenin (DIG)-tagged anti-sense probe hybridized to a target transcript is detected by anti-DIG antibodies conjugated with alkaline phosphatase enzyme (AP) (anti-DIG-AP). The hybrid molecules are visualized through an AP-catalyzed color reaction using as the substrate 5-bromo-4-chloro-3-indolyl phosphate (BCIP) and nitro blue tetrazolium chloride (NBT). This method can be applied to both free-living pre-parasitic juveniles and early endoparasitic stages of cyst nematodes.