EXPRESSION CORE

表达核心将确定项目I、II和III确定的候选基因表达的发育时间和组织。这些研究将使用免疫组织化学或原位杂交（视情况而定）确定RNA和/或蛋白质表达模式。 此外，将使用RCAS-TVA系统对特别感兴趣的基因进行野生型和显性阴性蛋白的组织表达分析。膜片整体安装很难检查，但该核心的主任和工作人员已经从阿克曼博士那里学到了必要的技术。肺器官培养已经在实验室中使用了20年。RNAi技术在过去的8年里一直被PI使用。 与果蝇核心一起，RNAi技术将成为该计划项目的唯一可用技术。下面简要介绍Loscertales博士在分析CDH候选基因时经常使用的方案。

The Expression Core will determine the developmental times and tissues in which candidate genes identified by Projects I, II and III are expressed. These studies will determine both RNA and/or protein expression patterns using immunohistochemistry or in situ hybridiation, whichever is deemed appropriate. Further, genes of particular interest will undergo analysis using the RCAS-TVA system for tissue expression of both wildtype and dominant negative proteins. Diaphragm whole mounts are difficult to examine, but the director and staff of this core have learned the necessary techniques from Dr. Akerman. Lung organ culture has been in use in the lab for 20 years. RNAi technology has been used by the Pis for the past 8 years. Together with the Drosophila Core, RNAi technology will be uniquely available for the program project. Brief descriptions of the protocols that Dr. Loscertales uses routinely which will be instrumental in analyzing CDH candidate genes follow.