Evolutionary studies of mouse seminal fluid protein genes

描述（由申请人提供）：本高级博士后奖学金申请的目标是在三个方面对申请人进行教育：1)基因组学和2)蛋白质组学，包括实验室技术和此类研究所需的计算机工具，以及3)比较杂交区不同基因渗入的相对选择强度所需的统计方法。本建议的具体目的是：1。利用蛋白质组学和基因组学方法鉴定小鼠精液的蛋白质成分。基因组学方法将使用来自小鼠前列腺的est现有数据来识别候选基因，蛋白质组学方法将使用高通量LC-MS/MS直接识别小鼠精液成分；2. 对上述确定的小鼠精液蛋白的候选基因进行测序：a)在五种不同的啮齿动物物种中，通过中性理论的统计测试进行分析，以寻找这些基因的特定位点选择证据（阳性和阴性）；b)在野生来源的家养m.m.和肌肉m.m.种群中，寻找这些基因中固定或几乎固定的多态性，用于下面目标4中概述的杂交区研究；3. 通过询问进化相似的基因是否属于小鼠、人类和其他哺乳动物基因组的共系区域，确定精液蛋白的真正同源基因；4. 通过绘制特定精液蛋白基因在欧洲小家鼠杂交带的渗入情况来测试它们的选择：a)发现在两个亚种之间具有固定或几乎固定多态性的候选基因（即在Aim 2a的选择下）（上述Aim 2b）；b)比较精液蛋白基因与相对低选择基因（如许多核同工酶）和高选择基因（如X和Y染色体多态性）在杂交区里的渗入曲线。

DESCRIPTION (provided by applicant): The goal of this senior postdoctoral fellowship application is to educate the applicant in three areas: 1) genomics and 2) proteomics, including both the laboratory techniques and the in silico tools necessary for such studies, and 3) the statistical methodology necessary for comparing the relative strength of selection on different genes introgressing across a hybrid zone. The specific aims of this proposal are: 1. To identify the protein components of mouse seminal fluid using proteomic and genomic approaches. The genomic approach will use existing data on ESTs from mouse prostate to identify candidate genes and the proteomic approach will use high-throughput LC-MS/MS to directly identify components of mouse seminal fluid; 2. To sequence candidate genes for the mouse seminal fluid proteins identified above a) in five different rodent species for analysis by statistical tests of the Neutral Theory to look for site-specific evidence of selection (positive and negative) on these genes and b) in wild-derived M. m. domesticus and M. m. musculus populations to look for fixed or nearly fixed polymorphisms in these genes for use in hybrid zone studies outlined in Aim 4 below; 3. To identify true orthologous genes for seminal fluid proteins by asking whether evolutionary similar genes fall within syntenic regions of mouse, human, and other mammalian genomes; 4. To test for selection on specific seminal fluid protein genes by plotting their introgression across the Mus musculus hybrid zone in Europe: a) to discover candidate genes (i.e., shown to be under selection by Aim 2a) that have a fixed or nearly fixed polymorphism between the two subspecies (Aim 2b above); and b) to compare clines of introgression of seminal fluid protein genes across the hybrid zone for seminal fluid protein genes with clines for genes under relatively low selection (e.g., many nuclear allozymes) and genes under high selection (e.g., X and Y chromosome polymorphisms). Relevance: Seminal fluid proteins show dynamic evolutionary histories, significant positive selection and variable selective pressure between lineages. As our understanding of the roles of seminal fluid components grows, so will our understanding of factors that influence reproductive success in humans and other animals. This proposal will provide a comparative genomics view of mammalian seminal fluid proteins from rodents and primates, which should lead to a better understanding of, and ability to cope with human problems involving infertility. Furthermore, the major source of seminal fluid proteins is the prostate, a common site of male cancer. Disease research may benefit from studies of selection, since positive selection is often associated with human disease genes.