Evolutionary studies of mouse seminal fluid protein genes

描述（由申请人提供）：该高级博士后奖学金申请的目标是在三个领域教育申请人：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。该提案的具体目标是：1.利用蛋白质组学和基因组学方法鉴定小鼠精液中的蛋白质组分。基因组学方法将使用现有的小鼠前列腺EST数据来鉴定候选基因，蛋白质组学方法将使用高通量LC-MS/MS来直接鉴定小鼠精液的组分; 2.对上面鉴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学检验进行分析，以寻找这些基因上的位点特异性选择证据（阳性和阴性），和B）在野生来源的M. M. Acetylticus和M. M.小家鼠群体中寻找这些基因中固定或几乎固定的多态性，用于下文目标4中概述的杂交区研究; 3.通过询问进化上相似的基因是否属于小鼠、人类和其他哺乳动物基因组的同线区域，来鉴定精液蛋白的真正直链基因; 4.为了通过绘制特定精液蛋白基因在欧洲小家鼠杂交区的渐渗来测试对它们的选择：a）发现候选基因（即，显示在Aim 2a）的选择下，其在两个亚种之间具有固定或几乎固定的多态性（上述Aim 2 B）;和B）比较精液蛋白基因在杂交区中的精液蛋白基因渐渗的谱系与在相对低选择下的基因的谱系（例如，许多核等位酶）和高度选择下的基因（例如，X和Y染色体多态性）。 相关性：精液蛋白质具有动态的进化历史、显著的正选择和不同谱系间的选择压力。随着我们对精液成分作用的理解不断加深，我们对影响人类和其他动物生殖成功的因素的理解也将不断加深。这项建议将提供一个比较基因组学的观点哺乳动物精液蛋白从啮齿动物和灵长类动物，这应该会导致更好地理解，并有能力科普人类的问题，涉及不育。此外，精液蛋白质的主要来源是前列腺，这是男性癌症的常见部位。疾病研究可能会受益于选择的研究，因为积极的选择往往与人类疾病基因有关。

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.