Core--Functional Geomics and Bioinformatics

来自不同实验室的研究，包括SBRP研究人员的研究，已经确定在体内特定生物转化酶的水平与许多毒理学问题有关。例如，在几项分子流行病学调查中，已经确定了生物转化基因的特定多态与环境相关疾病的发病率之间的遗传相关性。此外，生物监测越来越多地涉及对个体和前哨动物的遗传/分子指数的评估，目的是检测疾病易感性的可能标记，以及确定化学效应的早期指标--例如，由于暴露于环境毒物，生物转化酶基因表达谱发生变化。技术支持方面的进展 分子研究导致了涉及这些生物转化酶系统的复杂技术的发展，这些技术适用于包括新的修复策略在内的广泛的实验问题。UW SBRP功能基因组学和生物信息学核心C实验室的长期目标是为UW SBRP研究人员提供服务设施，使其能够开发新型基因分析、高通量遗传和蛋白质组分析、最先进的定量基因表达和生物信息技术，并与环境相关疾病机制相关。所有六个单独的UW SBRP项目都将利用UW SBRP核心C实验室，并将受益于集中服务。这些核心工作将侧重于广泛的与分子生物学和蛋白质组相关的技术，这些技术可能会预测到：接触毒物；生理和神经功能受损；对环境中发生的毒剂，特别是危险废物场地中常见的毒剂造成的神经损害的异常敏感性。在这项提议的5年期间，华盛顿大学SBRP核心C实验室将提供以下服务：提取1,200个样本的DNA/RNA，分离625个单核细胞样本，对255个GeneChip和98个cDNA鲑鱼阵列进行处理和生物信息学分析，生产一个定制的代表多达2,500个的“SNP芯片” 遗传变异及随后的加工和遗传关联分析，鉴定多达约500万个基因类型，开发25个新的基于TaqMan的基因分型分析，构建131个新的基因表达分析，执行约7,800个基因表达分析，提供约4,000个DNA测序凝胶通道，产生10个多肽衍生抗体，并创建15个 新奇的ELISA。华盛顿大学SBRP核心C实验室提供的这一共享资源支持以具有成本效益和效率的方式实施必要的功能基因组分析，从而最大限度地为UW SBRP研究人员提供这些重要工具。

Research from various laboratories, including those of SBRP investigators, has established that in vivo levels of particular biotransformation enzymes are relevant to numerous toxicological issues. For example, genetic correlates have been identified in several molecular epidemiologic investigations between particular polymorphisms in biotransformation genes and the incidence of environmental-related disease. In addition, biological monitoring has increasingly involved the assessment of genetic/molecular indices from individuals and sentinel animals, for the purpose of detecting possible markers of disease susceptibility, as well as identification of early indicators of chemical effect - such as alterations in biotransformation enzyme gene expression profiles due to exposure to environmental toxicants. Advances in technology supporting molecular research have led to the development of complex techniques involving these biotransformation enzyme systems, which are applicable to a broad range of experimental problems including novel remediation strategies. The long term objective of the UW SBRP Functional Genomics and Bioinformatics Core C Laboratory is to provide a service facility for UW SBRP investigators enabling novel genetic assay development, high throughput genetic and proteomic analysis, state-of-the-art quantitative gene expression and bioinformatic technologies, and correlates to environmental-related disease mechanisms. All six individual UW SBRP projects will utilize the UW SBRP Core C Laboratory and will benefit from the centralized services. These Core efforts will focus on a wide range of molecular biology- and proteomic-related technologies that may be predictive of: exposure to toxicants; impaired physiologic and neurologic function; unusual susceptibility to neurological damage from toxic agents that occur in the environment, particularly those that are commonly present at hazardous waste sites. Over the 5 year duration of this proposal, the UW SBRP Core C Laboratory will provide the following services: DNA/RNA extraction of 1,200 samples, the isolation of 625 mononuclear cell samples, processing and bioinformatic analyses of 255 GeneChips and 98 cDNA salmonoid arrays, the production of a custom "SNP chip" representing up to 2,500 genetic variants and subsequent processing and genetic association analyses, the identification of up to approximately 5,000,000 genotypes, the development of 25 new TaqMan-based genotyping assays, the construction of 131 new gene expression assays, the execution of approximately 7,800 gene expression analyses, the provision of approximately 4,000 DNA sequencing gel lanes, the generation of 10 peptide derived antibodies, and the creation of 15 novel ELISAs. This shared resource provided by the UW SBRP Core C Laboratory supports implementation of the requisite functional genomic assays in a cost-effective and efficient manner, therefore maximizing the availability of these important tools for UW SBRP investigators.