GNOTOBIOTICS AND MICROBIOLOGY CORE

基因组学和微生物学核心的比活性 A.核心服务 核心服务以按存储容量使用计费的方式提供。它们旨在实现购买设备、试剂/用品和劳动力的规模经济，从而使个人调查人员的实际使用成本得到高达50%的补贴。 1)无菌小鼠的维护：核心为许多研究人员维护无菌小鼠，包括野生型瑞士韦伯斯特和C57BL/6菌株，以及由Taconic或其他网站为研究人员衍生的基因敲除菌株。叶利塞耶夫为4名定期使用岩心隔离器的调查人员管理隔离器。Belavusava女士协助通过培养和16S rRNA聚合酶链式反应对小鼠进行微生物无菌检查。 2)特定关联小鼠的维护：核心有4个小型正压隔离器，用于与共生生物进行特定关联实验。核心主任和叶利塞耶夫与调查人员合作，为特定关联实验设计实验计划，并安排隔离器的使用。 3)与BL-2病原体特异关联的小鼠的维护：核心有2个负压隔离器，用于将无菌小鼠单独与BL-2病原体关联或与共生物种关联。Yelisyev先生在Belavusava女士的帮助下，维护着与特定相关小鼠的隔离物，而Belavusava女士则协助培养用于联合的分离物，并对来自 被殖民的老鼠，这是理所当然的。 4)现场饲养和野生型无菌库的可获得性：核心维护2个大型繁殖隔离器，主要用于提供本地供应的无菌瑞士韦氏和C57BL/6小鼠，用于实验。 当地饲养为HDDC研究人员节省了大量的动物购买成本(节省70%)，而且通常提供比商业来源更容易获得的野生型小鼠。根据要求，该中心还将保留少量其他品系的无菌小鼠的繁殖对进口 由人类发展中心调查人员进行指定实验。耶利塞耶夫维持着动物繁育群体，包括订购商业饲养的无菌小鼠，以根据需要补充繁育种群。 注意，鉴于朗伍德地区可用鼠标设施的空间限制，核心E目前不进行无菌衍生。核心确实提供了信息和选项，通过Taconic或Charles River将小鼠品系衍生到无菌状态。 首页--期刊主要分类--期刊细介绍--期刊题录与文摘--期刊详细文摘内容 5)组织收获和组织病理学：帮助收集小鼠组织和血液，并为冰冻切片或石蜡包埋准备材料。耶利塞耶夫精通动物解剖，可以进行组织采集或以其他方式协助调查人员。Bry博士还能够为下游应用提供最佳的老鼠身体解剖方法并提供建议。 6)好氧和厌氧培养：核心提供好氧、微好氧和厌氧物种培养的设备、材料和专业知识。Bry博士和Onderdonk博士的实验室提供了资源，并可能通过他们实验室的额外技术人员提供进一步的支持。 7)微生物鉴定和分析：核心拥有通过微生物、生物化学和分子方法鉴定共生菌株的设备、资源和专业知识，包括发酵图谱和其他酶活性的生化分析，短、中、长链脂肪酸的MIDI分析，以及16S rRNA序列与精选的16S文库的比较。 8)微生物组分析。该中心拥有资源和专业知识，协助研究人员收集和准备人类和啮齿动物样本，以便对16S rRNA基因签名进行深度测序。 HDDC调查人员可以访问BWH高性能计算集群上维护的生物信息学工具，以分析16S rRNA基因签名的罗氏454测序数据。

SPECIFIC ACTIVITIES OF THE GNOTOBIOTICS and MICROBIOLOGY CORE A. Core Services Core services are provided on a charge-back basis. They are designed to achieve economies of scale for purchase of equipment, reagents/supplies, and labor so that the real costs of use are subsidized by up to 50% for individual investigators. 1) Maintenance of Germfree mice: The core maintains germfree mice for a number of investigators, including wild-type Swiss Webster and C57BL/6 strains, as well as knockout strains that have been derived for investigators by Taconic or other sites. Mr. Yeliseyev manages isolators for 4 investigators who use core isolators on a regular basis. Ms. Belavusava assists in microbiological sterility checks of mice, as performed by culture and 16S rRNA PCR. 2) Maintenance of Specifically-Associated Mice: The core has 4 small positive-pressure isolators for specific-association experiments with commensal organisms. The Core Director and Mr. Yeliseyev work with investigators to devise experimental plans for specific-association experiments and schedule use of isolators. 3) Maintenance of Specifically-Associated Mice with BL-2 pathogens: The core has 2 negative-pressure isolators for association of germfree mice with BL-2 pathogens alone or with commensal species. Mr. Yeliseyev, with assistance from Ms. Belavusava, maintains isolators with specifically associated mice while Ms. Belavusava assists with culturing of isolates for associations and quantitative culture of materials from colonized mice, as warranted. 4) On-site breeding and availability of wild-type germfree stocks: The core maintains 2 large breeding isolators, primarily to provide a local supply of germfree Swiss-Webster and C57BL/6 mice for experiments. Local breeding provides a significant savings in animal purchase costs to HDDC investigators (>70% savings) and often provides a more ready supply of wild-type mice than can be obtained from commercial sources. Per request, the core will also maintain small numbers of breeding pairs of other strains of germfree mice imported by HDDC investigators for designated experiments. Mr Yeliseyev maintains animal breeding colonies, including ordering commercially-maintained germfree mice to replenish breeding stocks as needed. Note, Core E does not currently undertaking germfree derivations given space constraints in available mouse facilities in the Longwood area. The core does provide information and options for deriving mouse lines to a germfree state through Taconic or Charles River. HDDC: Integrated Epithelial and Mucosal Biology Lencer, Wayne I. 5) Tissue harvest and histopathology: The core offers assistance with collection of mouse tissues and blood, and preparation of materials for frozen section or paraffin embedding. Mr. Yeliseyev is well-versed in animal dissection and is available to perform or otherwise assist investigators with tissue harvests. Dr. Bry is also able to assist and advise with optimal methods to perform mouse necropsies for downstream applications. 6) Aerobic and anaerobic culture: The core provides equipment, materials and expertise for the culture of aerobic, microaerophilic and anaerobic species. Resources are available in Dr. Bry and Dr. Onderdonk's labs and may be further supported through additional technical staff in their labs. 7) Microbial identification and analysis: The core has equipment, resources and expertise to speciate commensal isolates by microbiological, biochemical and molecular methods, including biochemical analyses of fermentation profiles and other enzyme activities, MIDI analysis of short- medium- or long-chain fatty acids, and comparison of 16S rRNA sequences against curated 16S libraries. 8) Analyses of the Microbiome. The core has the resources and expertise to assist investigators with the collection and preparation of human and rodent samples for deep sequencing of 16S rRNA gene signatures. HDDC investigators may access bioinformatics tools maintained on the BWH High Performance Computing Cluster for analysis of Roche 454 sequencing data of 16S rRNA gene signatures.