Molecular Biology Shared Resource

分子生物学共享资源

• 批准号: 7698816
• 负责人: SHIRLEY M TAYLOR
• 金额: $ 1.02万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-03 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7698816
• 关键词: Animals; Biochemistry; Biology; Cancer Center; Characteristics; Chemicals; Chemistry; Client; Cloning; Collaborations; Communities; Complementary DNA; Consultations; Core Facility; Custom; Daily; Endotoxins; Epitopes; Equipment and supply inventories; Event; Facility Construction Funding Category; Faculty; Feedback; Funding; Gene Targeting; Generations; Genomic Library; Grant; Head; Human Resources; Individual; Insecta; Knock-out; Knockout Mice; Laboratories; Laboratory Personnel; Laboratory Technicians; Libraries; Maintenance; Malignant Neoplasms; Mammalian Cell; Maps; Methods; Mission; Molecular; Molecular Analysis; Molecular Biology Shared Resource; Occupations; Peer Review; Phase; Plasmid Cloning Vector; Plasmids; Polymerase Chain Reaction; Postdoctoral Fellow; Procedures; Production; Proteins; Published Comment; Quality Control; Range; Recording of previous events; Refractory; Research; Research Personnel; Research Project Grants; Research Support; Resource Sharing; Resources; Respondent; Screening procedure; Services; Site; Students; Support of Research; Surveys; System; Techniques; Therapeutic; Time; Training; Transgenic Organisms; Update; Variant; Vendor; Virus; Work; base; cost; design; design and construction; embryonic stem cell; experience; expression vector; homologous recombination; medical specialties; member; new technology; programs; protein expression; response; scale up; user-friendly; vector

The construction and manipulation of plasmids and of cloned DMA sequences is a central activity of every laboratory involved in the molecular analysis of cancer. Some manipulations are more challenging and some plasmids refractory to common procedures. The Molecular Biology Shared Resource was initiated to assist Cancer Center investigators with plasmid manipulations and cloning projects that prove problematic to individual laboratories or that prove refractory to manipulation by laboratory technicians, students, and post-docs. In addition, the Molecular Biology Shared Resource provides a cost-effective alternative to a variety of basic services to the investigators of the Massey Cancer Center (MCC). These basic services include endotoxin-free plasmid purification, insert purification, plasmid management and maintaining an inventory of plasmid vectors and clones. Other projects are designed upon the specific needs of client laboratories including custom subcloning, library screening, generation and optimization of protein expression vectors, design, construction, and analysis of gene targeting vectors, and protein production in bacterial, insect and mammalian cells. The MBSR works in synch with the Transgenic and Knockout Core Facility and facilitates access of Cancer Center members to such animals through vector design and construction. A large part of the activity of the Core involves plasmid manipulations that have proven difficult for the requesting laboratory, so each job tends to have unique characteristics and usually involves trouble-shooting and redesign of procedures. Thus, the expertise of Facility personnel has proven to be one of the most valuable services offered by the Core. MSBR personnel offer training seminars and previews of newer technologies as they are developed, and often train laboratory personnel in routine (and not so routine) molecular procedures. The Core Director offers a formal course in all phases of molecular technique that can be taken for credit or audited. New directions for the Molecular Biology Shared Resource are continuously being sought, in consultation with Cancer Center investigators and the MBSR oversight committee. The recent hire of new faculty in Chemical Biology into the Departments of Chemistry and Biochemistry (members of the Molecular Cancer Therapeutics program) has promoted a move towards the support of protein production for structural studies. The Shared Resource has been increasingly active during the past 2 years assisting investigators with their protein production needs, generating expression vectors and optimizing protein expression systems for scaled-up production.

质粒和克隆的DNA序列的构建和操作是本发明的核心活动。 所有参与癌症分子分析的实验室有些操作更具挑战性 和一些质粒对常规方法是难治的。分子生物学共享资源 开始协助癌症中心的研究人员进行质粒操作和克隆项目， 对个别实验室有问题或实验室技术人员难以操作， 学生和博士后。此外，分子生物学共享资源提供了一个具有成本效益的 梅西癌症中心（MCC）的研究人员的各种基本服务的替代品。这些 基本服务包括无内毒素质粒纯化、插入物纯化、质粒管理和 维持质粒载体和克隆的库存。其他项目是根据具体的 客户实验室的需求，包括定制亚克隆、文库筛选、 蛋白质表达载体，基因靶向载体的设计、构建和分析，以及蛋白质 在细菌、昆虫和哺乳动物细胞中生产。正念减压疗法与转基因同步工作， 敲除核心设施，并促进癌症中心成员通过载体接触这些动物 设计和施工。核心蛋白的大部分活性涉及质粒操作， 事实证明，对于请求实验室来说很难，因此每项工作往往都有独特的特点， 包括解决问题和重新设计程序。因此，设施人员的专业知识已经证明， 成为内核提供的最有价值的服务之一MSBR人员提供培训研讨会， 预览新技术的发展，并经常培训实验室人员在日常（和 不那么常规）分子程序。核心主任提供了一个正式的课程，在所有阶段的分子 可以被认为是信用或审计的技术。 分子生物学共享资源的新方向正在不断寻求， 与癌症中心研究人员和正念减压疗法监督委员会进行磋商。最近雇用的新 化学生物学教师进入化学和生物化学系（分子 癌症治疗计划）已经推动了一项支持蛋白质生产的举措， 问题研究在过去两年中，共享资源越来越积极地协助调查人员 根据他们的蛋白生产需求，生成表达载体并优化蛋白表达 扩大生产的系统。