Expression, Biochemistry and Immune Reagent Core

表达、生化和免疫试剂核心

• 批准号: 8687588
• 负责人: Mark T Heise
• 金额: $ 39.3万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8687588
• 关键词: Affect; Alphavirus; Animals; Antibody Formation; Antigens; Antiviral Agents; Antiviral Response; Antiviral Therapy; Apoptosis; Biochemical; Biochemistry; Biological Assay; Biology; Cells; Characteristics; Collaborations; Coronavirus; DNA Viruses; Development; Ebola virus; Ensure; Filovirus; Functional RNA; Genes; Genetic Transcription; Goals; Human; Human Herpesvirus 8; Immune; Immune Sera; Immune response; Infection; Influenza A virus; Interferons; Lentivirus Vector; Medical; Molecular Virology; Monoclonal Antibodies; Mus; Open Reading Frames; Pathogenesis; Pathway interactions; Post-Translational Protein Processing; Process; Production; Proteins; Public Health; Quality Control; RNA; RNA Viruses; Reagent; Replication-Associated Process; Replicon; Research Personnel; Research Project Grants; SARS coronavirus; Signal Transduction; Specificity; Subfamily lentivirinae; System; Technology; Therapeutic Intervention; Untranslated RNA; Vaccinated; Viral; Viral Pathogenesis; Viral Proteins; Virus; Virus Diseases; Work; base; expression vector; glycosylation; influenzavirus; insight; meetings; novel; novel vaccines; particle; pathogen; programs; protein expression; protein function; protein protein interaction; response; vector; vector vaccine; viral DNA

Emerging pathogens, such as SARS-CoV, Emerging Coronavirus 2012 {EMC/2012), influenza virus (lAV), Filoviruses (Ebola) and Human Herpesvirus 8 {HHV8) are significant public health threats. All of these pathogens encode uncharacterized open reading frames (ORFs) and noncoding RNAs that we hypothesize will act as either positive or negative regulators of viral replication or modulators of host cell regulatory processes or antiviral responses. Therefore, investigating how these ORFs/RNAs affect viral replication and/or the host response will enhance our general understanding of the pathogenesis of each of these viruses and may identify common pathways that can be targeted for therapeutic intervention. Our program involves a highly interactive group of investigators, who will work collaboratively to characterize the impact of novel viral ORFs and/or noncoding RNAs in regulating viral replication or key host processes that are likely to be targets of viral modulation, such as type I IFN response, apoptosis, or inflammasome activation. This analysis requires that each research project has access to a standardized set of expression systems that allow for ready cross comparison of protein/RNA function between projects. Therefore, the core will clone and express candidate ORFs/noncoding RNAs from standard sets of expression vectors, perform quality control assays to ensure appropriate expression of the candidate ORF/RNA, and then provide these expression vectors to each of the research projects. Since one of the goals of this program is to characterize how novel ORFs affect viral pathogenesis, we will generate antisera against each of the candidate ORFs, validate the specificity of this antiserum, and then provide it to each project for use in assessing protein expression, cellular localization, and protein/protein interactions in the context of authentic SARS-CoV, EMC/2012, lAV, Ebola virus, or HHV 8 infection. Lastly, for each candidate ORF, the core will use the expression vectors to perform detailed analysis of the expression and biochemical characteristics of each ORF (e.g. posttranslational modifications such as glycosylation) in collaboration with each research project.

新兴病原体，例如SARS-COV，新兴冠状病毒2012（EMC/2012），流感病毒（LAV）， 丝状病毒（埃博拉病毒）和人类疱疹病毒8（HHV8）是重大的公共卫生威胁。所有这些 病原体编码我们假设的未表征的开放式阅读框（ORF）和非编码RNA 将充当病毒复制或宿主细胞调节调节剂的正调节剂或负调节剂 过程或抗病毒反应。因此，研究这些ORF/RNA如何影响病毒复制 和/或宿主反应将增强我们对每种发病机理的一般理解 病毒，可以识别可以针对治疗干预的常见途径。 我们的计划涉及一个高度互动的调查人员，他们将共同努力以表征 新型病毒ORF和/或非编码RNA在调节病毒复制或关键宿主过程中的影响 可能是病毒调节的靶标，例如I型IFN反应，凋亡或炎性体 激活。该分析要求每个研究项目都可以访问一套标准化的表达式 可以在项目之间进行蛋白质/RNA功能的即时比较的系统。因此， 核心将克隆和表达候选ORF/非编码RNA来自标准表达向量集， 执行质量控制测定，以确保候选ORF/RNA的适当表达，然后 向每个研究项目提供这些表达向量。由于该程序的目标之一是 为了表征新颖的ORF如何影响病毒发病机理，我们将对每一个产生抗血清 候选ORF，验证该抗血清的特异性，然后将其提供给每个项目以供在 评估蛋白质表达，细胞定位和蛋白质/蛋白质相互作用 SARS-COV，EMC/2012，LAV，埃博拉病毒或HHV 8感染。最后，对于每个候选人，核心将 使用表达向量对表达和生化特征进行详细分析 每种ORF（例如翻译后修饰，例如糖基化） 项目。