Cogswell Alexis A, Collins James J, Newmark Phillip A, Williams David L
Rush University Medical Center, Department of Immunology Microbiology, Chicago, IL, United States.
Mol Biochem Parasitol. 2011 Jul-Aug;178(1-2):46-50. doi: 10.1016/j.molbiopara.2011.03.001. Epub 2011 Mar 21.
The genome sequence for Schistosoma mansoni has been determined, allowing the complete protein complement to be predicted. However, few functional genomics techniques have been developed for use in S. mansoni, limiting the usefulness of the sequence data. Here we describe a whole mount in situ hybridization (WISH) method that can be used to identify the tissue-specific expression of transcripts in S. mansoni. Using this protocol we determine the tissue-specific expression of tetraspanin 2, a female-enriched tetraspanin, phenol oxidase, the secretory Cu/Zn superoxide dismutase, and an Argonaute family member. The localization of these transcripts by WISH correlates with prior studies performed using immunohistochemistry and/or in situ hybridization on tissue sections. WISH can be adapted to screen multiple transcripts, thus identifying novel targets for drugs or vaccines.
曼氏血吸虫的基因组序列已被测定，这使得完整的蛋白质组得以预测。然而，针对曼氏血吸虫开发的功能基因组学技术很少，限制了序列数据的实用性。在此，我们描述了一种全组织原位杂交（WISH）方法，该方法可用于鉴定曼氏血吸虫中转录本的组织特异性表达。使用该方案，我们确定了四跨膜蛋白2（一种在雌性中富集的四跨膜蛋白）、酚氧化酶、分泌型铜/锌超氧化物歧化酶和一个AGO蛋白家族成员的组织特异性表达。通过WISH对这些转录本的定位与先前使用免疫组织化学和/或组织切片原位杂交进行的研究结果相关。WISH可用于筛选多个转录本，从而确定药物或疫苗的新靶点。
