Transcriptional Regulation of Oocyte-Specific Genes

描述(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可受精卵，这些细胞器和结构在受精时具有独特的功能。本研究的目的是通过检测卵母细胞特异性基因的转录调控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有基因的协调表达，其中一些调节因子将对卵母细胞具有选择性。为了验证这一假设，我将检测几个卵母细胞特有基因启动子中的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地丰富。计算算法将被用来识别卵母细胞基因启动子中的保守顺式元件，随后将使用报告基因结构在体内测试转录活性。我们将通过计算、突变分析、生物化学和分子遗传学的方法来鉴定顺式元件中具有假定结合位点的反式作用因子。定义调控几个卵母细胞基因启动子的转录复合体将在基因调控网络中创建一个范例，该网络协调卵母细胞的发育，并促进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.