Regulated expression of B. burgdorferi virulence genes

伯氏疏螺旋体毒力基因的调节表达

• 批准号: 7936229
• 负责人: Felipe Cardenas Cabello
• 金额: $ 38.28万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7936229
• 关键词: Adhesions; Antibiotics; Antisense RNA; Bacteria; Bacterial Adhesins; Binding; Biological Assay; Biological Process; Biology; Borrelia; Borrelia burgdorferi; Collagen; Collagen Type I; Complement; Data; Detection; Development; Extracellular Matrix; Gene Expression; Gene Fusion; Genes; Genetic; Genome; Genomics; Goals; Grant; Hybrids; In Vitro; Individual; Knockout Mice; Latex; Libraries; Lyme Disease; Mediating; Methodology; Microbial Genetics; Molecular; Molecular Genetics; Mus; Order Spirochaetales; Organism; Output; Pattern; Plasmids; Population; Preparation; Property; RNA library; Research; Rest; Role; Route; Screening procedure; Skin; System; Tetanus Helper Peptide; United States; Vector-transmitted infectious disease; Virulence; Work; base; design; gene function; genetic manipulation; in vivo; mutant; novel; novel vaccines; pathogen; promoter; public health relevance; tool

DESCRIPTION (provided by applicant): The ability to extract biologically relevant information from the genome of Borrelia burgdorferi, the cause of Lyme disease, continues to be hampered by the lack of genetic tools to identify the biological functions of the many genes of unknown functions that it contains. We and others have adapted a number of classical microbial genetics tools to B. burgdorferi and have used them to isolate null mutants and their complemented derivatives in order to analyze the contribution of these genes to B. burgdorferi biology and virulence. No one yet has been able to produce genomic saturated libraries or to isolate conditional and conditional lethal mutants of this pathogen. The long-term goal of our research is the identification and characterization of virulence determinants of B. burgdorferi. One group of potential virulence determinants that may be involved in persistence of this organism in mammalian hosts is the adhesins and adhesin regulators mediating its adhesion to Type I collagen in the extracellular matrix. Data obtained by us during the previous project period indicated that gene expression in B. burgdorferi could be modulated in a regulated fashion in a B. burgdorferi strain constitutively expressing the Tet repressor using modified B. burgdorferi hybrid promoters containing tetO operators. This system was used with the non-antibiotic inducer anhydrotetracycline (ATc) to regulate gene expression by gene fusions and antisense RNA, and in preparing and screening antisense RNA libraries. We also found that the mariner transposon system could be used to isolate transposition mutants in B. burgdorferi chromosomal and plasmid genes. On the basis of a combination of these findings, we propose the following Specific Aims. 1) Identify and characterize novel B. burgdorferi Type 1 collagen adhesins/adhesin regulators using antisense RNA libraries, isogenic mutants for adhesins/adhesins regulators, and microarrays. 2) Characterize B. burgdorferi genes relevant to colonization of the extracellular matrix in chronically infected MyD88 knockout mice using a global genomic approach involving saturated B. burgdorferi genomic transposon libraries screened by microarrays. 3) Complete development of novel systems of ATc - regulated gene expression in B. burgdorferi to study its adhesion to the ECM. PUBLIC HEALTH RELEVANCE: Lyme disease is the most common vector-borne disease in the United States. This project will develop new molecular genetic tools for studying the function of the genes of the Lyme disease bacterium, B. burgdorferi. These tools will be used to identify genes that are potentially responsible for this organism's virulence and which could be targets for new vaccines and antibiotics against Lyme disease.

描述（由申请人提供）：由于缺乏遗传工具来鉴定其包含的许多功能未知的基因的生物学功能，从伯氏疏螺旋体（莱姆病的病因）的基因组中提取生物学相关信息的能力仍然受到阻碍。我们和其他人将许多经典的微生物遗传学工具应用于伯氏疏螺旋体，并使用它们分离无效突变体及其互补衍生物，以分析这些基因对伯氏疏螺旋体生物学和毒力的贡献。目前还没有人能够产生基因组饱和文库或分离这种病原体的条件和条件致死突变体。我们研究的长期目标是鉴定和表征伯氏疏螺旋体的毒力决定因素。可能与该生物体在哺乳动物宿主中的持久性有关的一组潜在毒力决定因素是介导其与细胞外基质中的 I 型胶原的粘附的粘附素和粘附素调节剂。我们在上一个项目期间获得的数据表明，使用含有 tetO 操纵子的改良伯氏疏螺旋体杂交启动子，可以在组成型表达 Tet 阻遏物的伯氏疏螺旋体菌株中以受调控的方式调节伯氏疏螺旋体中的基因表达。该系统与非抗生素诱导剂脱水四环素（ATc）一起使用，通过基因融合和反义RNA调节基因表达，并用于制备和筛选反义RNA文库。我们还发现，mariner 转座子系统可用于分离伯氏疏螺旋体染色体和质粒基因中的转座突变体。根据这些发现的结合，我们提出以下具体目标。 1) 使用反义 RNA 文库、粘附素/粘附素调节剂的同基因突变体和微阵列来鉴定和表征新型伯氏疏螺旋体 1 型胶原粘附素/粘附素调节剂。 2) 使用涉及通过微阵列筛选的饱和伯氏疏螺旋体基因组转座子文库的全局基因组方法，表征与慢性感染的 MyD88 敲除小鼠中细胞外基质定植相关的伯氏疏螺旋体基因。 3) 完成新的 ATc 系统的开发 - 调节伯氏疏螺旋体的基因表达，以研究其对 ECM 的粘附。公共卫生相关性：莱姆病是美国最常见的媒介传播疾病。该项目将开发新的分子遗传学工具，用于研究莱姆病细菌伯氏疏螺旋体的基因功能。这些工具将用于识别可能导致该生物体毒力的基因，并可能成为针对莱姆病的新疫苗和抗生素的目标。

项目成果

Erythromycin resistance in Borrelia burgdorferi.

伯氏疏螺旋体对红霉素的耐药性。

• DOI: 10.1128/aac.46.11.3637-3640.2002
• 发表时间: 2002
• 期刊: Antimicrobial agents and chemotherapy
• 影响因子: 4.9
• 作者: Terekhova,Darya;Sartakova,MarinaL;Wormser,GaryP;Schwartz,Ira;Cabello,FelipeC
• 通讯作者: Cabello,FelipeC

Borrelia chilensis, a new member of the Borrelia burgdorferi sensu lato complex that extends the range of this genospecies in the Southern Hemisphere.

• DOI: 10.1111/1462-2920.12310
• 发表时间: 2014-04
• 期刊: Environmental microbiology
• 影响因子: 5.1
• 作者: Ivanova LB;Tomova A;González-Acuña D;Murúa R;Moreno CX;Hernández C;Cabello J;Cabello C;Daniels TJ;Godfrey HP;Cabello FC
• 通讯作者: Cabello FC

Genome Sequence of Borrelia chilensis VA1, a South American Member of the Lyme Borreliosis Group.

• DOI: 10.1128/genomea.01535-14
• 发表时间: 2015-02-12
• 期刊: Genome announcements
• 作者: Huang W;Ojaimi C;Fallon JT;Travisany D;Maass A;Ivanova L;Tomova A;González-Acuña D;Godfrey HP;Cabello FC
• 通讯作者: Cabello FC

Sleeper cells: the stringent response and persistence in the Borreliella (Borrelia) burgdorferi enzootic cycle.

• DOI: 10.1111/1462-2920.13897
• 发表时间: 2017-10
• 期刊: Environmental microbiology
• 影响因子: 5.1
• 作者: Cabello FC;Godfrey HP;Bugrysheva JV;Newman SA
• 通讯作者: Newman SA

Localization of BmpA on the exposed outer membrane of Borrelia burgdorferi by monospecific anti-recombinant BmpA rabbit antibodies.

通过单特异性抗重组 BmpA 兔抗体将 BmpA 定位在伯氏疏螺旋体暴露的外膜上。

• DOI: 10.1128/iai.72.4.2280-2287.2004
• 发表时间: 2004
• 期刊: Infection and immunity
• 影响因子: 3.1
• 作者: Shin,JungheeJ;Bryksin,AntonV;Godfrey,HenryP;Cabello,FelipeC
• 通讯作者: Cabello,FelipeC