Core B: Molecular Biology, Protein Expression and Structural Biology
核心 B:分子生物学、蛋白质表达和结构生物学
基本信息
- 批准号:10175002
- 负责人:
- 金额:$ 65.52万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2018
- 资助国家:美国
- 起止时间:2018-09-01 至 2023-06-30
- 项目状态:已结题
- 来源:
- 关键词:AcidsAffinityAreaAwarenessBiologyCell LineChemistryChromatographyCoagulation ProcessCollaborationsCollectionContractsCore FacilityCrystallizationDataData AnalysesData CollectionDeuteriumDevelopmentDockingEnsureFactor VFactor VIIIFeasibility StudiesGoalsHemostatic functionHomeHuman ResourcesHydrogenIndividualLaboratoriesMammalian CellMeasurementMeasuresModelingMolecularMolecular BiologyMotivationPhysiologyPlasma ProteinsPost-Translational Protein ProcessingPreparationProductionPropertyProteinsProtocols documentationQuality ControlReactionReagentRecombinant ProteinsRecombinantsResearch PersonnelResolutionResourcesRoentgen RaysRunningScheduleSeriesSerumServicesSourceStructureSynchrotronsThrombosisTrainingUniversitiesVariantVisitVitamin KWorkX ray diffraction analysisX-Ray CrystallographyYeastsbasedata modelingexperienceexpression vectorhigh end computerinstrumentinterestliquid chromatography mass spectrometrymembermodel buildingnovelparallel computerprogramsprotein expressionprotein purificationrepositoryskillsstable cell linestructural biologysuccess
项目摘要
Core B Abstract
The molecular biology, protein expression and structural biology core is organized around the pooling of the
resources and expertise of the project leaders in the program. The conduct of this core is based on a model
that has been successfully implemented in the past 10 years and has been extremely productive. This
centralized resource provides service and assistance in molecular biology, bacterial/yeast expression and in
the preparation of stable mammalian cell lines over-expressing vitamin K-dependent proteins, factor V variants
and factor VIII variants. The core provides the facilities, hardware and assistance for large-scale protein
production runs with stable or transiently transfected mammalian cells in serum-free conditioned media. The
core will provide facilities for large scale isolation of recombinant proteins, the appropriate chromatography
strategies and will maintain the affinity columns necessary for this activity. The core will also provide some
quality control services particularly the quantitation of -carboxyglutamic acid content to verify the integrity of
expressed vitamin K-dependent proteins. Assistance and facilities will also be provided for the large-scale
isolation of coagulation proteins from plasma. Core B personnel will assist program members in implementing
sparse matrix crystal screens, liaise with the UPenn x-ray facility to allow the collection of diffraction data on
the home source and provide expertise for optimization of crystallization conditions with expanded screens.
The core supports high performance computers equipped with necessary programs for processing diffraction
data, model building by molecular replacement and refinement. The core will also provide support for small
angle x-ray scattering measurements on the Home source and data analysis. Hydrogen/deuterium exchange
studies will be performed in collaboration with Renhao Li and facilitated by a modest sub-contract to Emory
University. The Director will be responsible for the oversight and efficient conduct of these activities and for
ensuring that equitable service is provided to the projects
核心B摘要
分子生物学,蛋白质表达和结构生物学核心是围绕汇集的
项目负责人在项目中的资源和专业知识。这个核心的行为是基于一个模型
过去十年来,这一倡议得到了成功实施,并且非常富有成效。这
集中资源提供分子生物学、细菌/酵母表达和
过表达维生素K依赖性蛋白、因子V变体的稳定哺乳动物细胞系的制备
和因子VIII变体。核心为大规模蛋白质生产提供设施、硬件和辅助
在无血清条件培养基中用稳定或瞬时转染的哺乳动物细胞进行生产。的
核心将提供大规模分离重组蛋白的设施,适当的色谱
策略,并将维护此活动所需的亲和列。核心还将提供一些
质量控制服务,特别是定量β-羧基谷氨酸含量,以验证完整性
表达维生素K依赖蛋白。还将为大规模的
从血浆中分离凝固蛋白。核心B人员将协助项目成员实施
稀疏矩阵晶体屏幕,联络与宾夕法尼亚大学x射线设施,使衍射数据的收集
提供专业知识,用于使用扩展筛优化结晶条件。
核心支持高性能计算机配备必要的程序处理衍射
数据,通过分子置换和细化建立模型。核心还将为小型
关于X射线角散射测量的源程序和数据分析。氢/氘交换
研究将与李仁浩合作进行,并通过与埃默里大学的适度分包合同来促进
大学主任将负责监督和有效开展这些活动,并
确保为项目提供公平的服务
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Sriram Krishnaswamy其他文献
Sriram Krishnaswamy的其他文献
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{{ truncateString('Sriram Krishnaswamy', 18)}}的其他基金
Hemostasis and Thrombosis: Chemistry, Biology and Physiology
止血和血栓形成:化学、生物学和生理学
- 批准号:
9982398 - 财政年份:2018
- 资助金额:
$ 65.52万 - 项目类别:
Hemostasis and Thrombosis: Chemistry, Biology and Physiology
止血和血栓形成:化学、生物学和生理学
- 批准号:
9769852 - 财政年份:2018
- 资助金额:
$ 65.52万 - 项目类别:
Hemostasis and Thrombosis: Chemistry, Biology and Physiology
止血和血栓形成:化学、生物学和生理学
- 批准号:
10439604 - 财政年份:2018
- 资助金额:
$ 65.52万 - 项目类别:
Hemostasis and Thrombosis: Chemistry, Biology and Physiology
止血和血栓形成:化学、生物学和生理学
- 批准号:
10175000 - 财政年份:2018
- 资助金额:
$ 65.52万 - 项目类别:
Core B: Molecular Biology, Protein Expression and Structural Biology
核心 B:分子生物学、蛋白质表达和结构生物学
- 批准号:
10439607 - 财政年份:2018
- 资助金额:
$ 65.52万 - 项目类别:
2012 Hemostasis Gordon Research Conference and Gordon Research Seminar
2012止血戈登研究会议暨戈登研究研讨会
- 批准号:
8388646 - 财政年份:2012
- 资助金额:
$ 65.52万 - 项目类别:
Proteinase Allostery and the Regulation of Blood Coagulation
蛋白酶变构和凝血调节
- 批准号:
8463606 - 财政年份:2011
- 资助金额:
$ 65.52万 - 项目类别:
Proteinase Allostery and the Regulation of Blood Coagulation
蛋白酶变构和凝血调节
- 批准号:
8299448 - 财政年份:2011
- 资助金额:
$ 65.52万 - 项目类别:
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