Genetic Determinants of the hVISA Mechanism

hVISA 机制的遗传决定因素

基本信息

批准号：
7881677
负责人：
JOHN E GUSTAFSON
金额：
$ 29.24万
依托单位：
NEW MEXICO STATE UNIVERSITY LAS CRUCES
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-15 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7881677
关键词：
Antibiotic Resistance Antibiotics Antimicrobial Resistance Bioinformatics Cause of Death Cessation of life Clinical Computer software Data Data Set Databases Development Exhibits Foundations Funding Gene Expression Profile Gene Mutation Genetic Determinism Genome Genomics Genus staphylococcus Grant Growth Hospitals Infection Institution Instruction Investments Journals Manuscripts Microarray Analysis Minority Mission Mutation Nature Nosocomial Infections Patients Predisposition Process Reporting Research Research Activity Resistance Sequence Analysis Source Staphylococcus aureus Students Surface System Techniques Time Training Treatment Failure Underrepresented Minority United States National Institutes of Health Vancomycin base comparative genomics design flexibility methicillin resistant Staphylococcus aureus microbial novel pathogen programs tool transcriptomics

项目摘要

DESCRIPTION (provided by applicant): Genetic alterations leading to hetero-vancomycin intermediate (hVISA) expression presently defies characterization. A major problem in hVISA mechanistic studies has been the lack of isogenic vancomycin-susceptible and hVISA, especially related or isogenic real-world clinical strains. We have now characterized a clonal hVISA and vancomycin-susceptible S. aureus clinical strain set. The specific aims of this proposal are to determine: (1) genomic alterations that occur in a clinical hVISA and clones of this strain expressing elevated susceptible vancomycin MICs; (2) transcriptome alterations that occur as a result of hVISA mechanism acquisition and genetic alterations leading to elevated vancomycin susceptible MICs; and (3) transcriptome alterations due to vancomycin induction that occur in hVISA and S. aureus strains expressing elevated vancomycin susceptible MICs. The first aim will be accomplished by either 454-based genomic sequencing of a temporally isolated clonal strain set that includes hVISA and non-hVISA expressing varied low-level and elevated susceptible vancomycin MICs, followed by genome annotation and comparison, or Nimblegene comparative genomic sequencing. During this process the PI will be trained on 454-sequencing as well as the genome annotation pipelines XGI, Alpheus, and the experimental GenVar. The second and third aims will be accomplished by comparing and contrasting the transcriptomes of a clonal set of hVISA, non-hVISA strains expressing elevated susceptible vancomycin MICs and non-hVISA strains expressing low-level susceptible vancomycin MICs, isolated following growth with and without vancomycin. These aims will also be enhanced by applying an investigative transcriptome tool referred to as the Staphylococcus microarray meta-database (SAMMD). During this process the PI will be trained on the current NIAID-PFGRC-TIGR-supported S. aureus microarrays version 4 and microarray analysis software. These research objectives are designed to produce enormous datasets that will make this SCORE-funded PI competitive for traditional NIH funding.

描述（由申请人提供）：导致异质范学霉素中间体（HVISA）表达目前违反表征的遗传改变。 HVISA机械研究中的一个主要问题是缺乏敏感性万古霉素易感和HVISA，尤其是相关或同基因的现实世界临床菌株。现在，我们已经表征了克隆HVISA和可易感的金黄色葡萄球菌临床应变组。该提议的具体目的是确定：（1）在临床HVISA和该菌株的克隆中发生的基因组改变，表达易感性万古霉素MIC；（2）由于HVISA机制的获取和遗传改变而导致的导致万古霉素敏感麦克风升高的转录组改变；（3）在HVISA和金黄色葡萄球菌菌株中发生的万古霉素诱导引起的转录组改变，表达了万古霉素易感麦克风的升高。第一个目标将通过454个基于暂时的克隆应变集的基因组测序来实现，该测序包括HVISA和非HVISA表达各种低级和升高的易感万古霉素MIC，然后进行基因组注释和比较，或进行基因组相比较，或鼻nimblegene比较基因组测序。在此过程中，PI将在454个测序以及基因组注释管道XGI，Alpheus和实验Genvar进行训练。第二和第三目的将通过比较和对比，并与表达易感性万古霉素MIC和非HVISA菌株表达升高的非HVISA菌株的克隆集合的转录组和表达低水平敏感的Vancomycin MIC的非HVISA菌株，该菌株分离出来，并在没有Vancomycin的情况下分离出来。这些目标还将通过应用称为葡萄球菌微阵列元数据库（SAMMD）（SAMMD）的调查转录组工具来增强。在此过程中，PI将在当前的NIAID-PFGRC-TOGRPORPORPORPORTAPTRATE的S. Aureus Microares版本4和微阵列分析软件上进行培训。这些研究目标旨在生产庞大的数据集，从而使这一分数资助的PI竞争传统的NIH资金竞争。