Transcriptional Regulation of Oocyte-Specific Genes

描述（由申请人提供）：卵母细胞经历一个漫长的发育过程。卵母细胞从原始生殖细胞发育成具有特殊细胞器和结构的受精卵，具有独特的受精功能。本研究的目的是通过检查卵母细胞特异性基因的转录控制来阐明卵母细胞发育程序的分子机制。我假设卵母细胞利用一个转录网络来控制卵母细胞特异性基因的协调表达，并且这些调节因子中的一些将对卵母细胞具有选择性。为了验证这一假设，我将检查几个卵母细胞特异性基因启动子内的c/s调控序列，例如那些编码独特的卵母细胞结构蛋白、酶和转录因子的基因，这些基因在卵母细胞中选择性富集。计算算法将用于鉴定卵母细胞基因启动子内保守的顺式元件，随后使用报告基因构建体进行体内转录活性测试。在顺式元件中具有假定结合位点的反式作用因子将通过计算、突变分析、生化和分子遗传学方法进行鉴定。定义调控几个卵母细胞基因启动子的转录复合物将为基因调控网络创造一个范例，从而协调卵母细胞的发育，并进一步提高我们在生殖生物学方面的知识。

DESCRIPTION (provided by applicant): An oocyte undergoes a prolonged developmental process. From a primordial germ cell, oocytes develop into fertilizable eggs with specialized organelles and structures that function uniquely at fertilization. The objective of this research is to elucidate the molecular mechanism of the oocyte developmental program by examining transcriptional controls of oocyte-specific genes. I hypothesize that oocytes utilize a transcriptional network that controls the coordinated expression of oocyte-specific genes and that some of these regulatory factors will be selective for oocytes. To test this hypothesis, I will examine c/s-regulatory sequences within promoters of several oocyte-specific genes, such as those encoding unique oocyte structural proteins, enzymes, and transcription factors that are selectively enriched in the oocyte. Computational algorithms will be used to identify conserved cis-elements within oocyte gene promoters, followed by in vivo testing of transcriptional activity using reporter gene constructs. Trans-acting factors with putative binding sites within cis-elements will be identified by computational, mutational analysis, biochemical, and molecular genetic approaches. Defining transcription complexes that regulate several oocyte gene promoters will create a paradigm in gene regulatory networks that orchestrates the development of oocytes and further our knowledge in reproductive biology.