GENETIC MECHANISMS IN TESTIS DIFFERENTIATION

描述(根据申请者的描述改编)：调查人员计划 为已知在睾丸中表达的基因制造犬类克隆 发展和定义这些的时间和空间表达模式 正常犬类胚胎性腺中的基因。具体目标是：1) 克隆犬SOX-9的完整编码序列。调查人员是 目前正在用人SOX-9筛选犬基因组DNA文库 与犬类DNA杂交。同源区域将被亚克隆并 以序列分析为特征。2)克隆犬基因组DNA 管理信息系统(AMH)、Wt-L和SF-1的序列他们将使用标准库 筛选方法和聚合酶链式反应(PCR)鉴定克隆 基因组DNA文库。同源区域将被亚克隆，它们的 通过序列分析确认了同源性。3)刻画时态 以及Sry、Sox 9、Mis、Wt-1和SF-1在正常组织中的空间表达模式 犬胚胎性腺的逆转录聚合酶链式反应和原位杂交 与犬类探针杂交。在这项研究结束时，他们将 准备研究这些基因的时间和空间表达模式 受影响胚胎中的基因，这将是理解 导致睾丸发育异常的遗传机制。

DESCRIPTION (Adapted from applicant's description): The investigators plan to produce canine clones for genes that are known to be expressed in testis development and define the temporal and spatial expression patterns of these genes in normal canine embryonic gonads. The specific aims are: 1) To clone the entire coding sequence of canine SOX-9. The investigators are currently screening a canine genomic DNA library with a human SOX-9 that hybridizes to canine DNA. Homologous regions will be subcloned and characterized by sequence analysis. 2) To clone canine genomic DNA sequences of MIS (Amh), Wt-l, and SF-1. They will use standard library screening methods and polymerase chain reaction (PCR) to identify clones in a genomic DNA library. Regions of homology will be subcloned and their identity confirmed by sequence analysis. 3) To characterize the temporal and spatial expression patterns of Sry, Sox 9, MIS, Wt-1 and SF-1 in normal canine embryonic gonads, using reverse transcriptase PCR and in situ hybridization with canine probes. At the end of this study, they will be poised to examine the temporal and spatial expression patterns of these genes in affected embryos, which will be critical to understanding the genetic mechanisms responsible for abnormal testis development.