MOLECULAR AND FUNCTIONAL STUDIES OF H INFLUENZAE B PILI

乙型流感病毒的分子和功能研究

• 批准号: 3139134
• 负责人: JANET R GILSDORF
• 金额: $ 15.9万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-01-01 至 1993-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3139134
• 关键词: Haemophilus influenzae; Macaca fascicularis; bacterial genetics; electron microscopy; gene expression; genetic manipulation; molecular pathology; mutant; pilus; transposon /insertion element; virulence

DESCRIPTION: (Adapted from applicant's abstract). The long range goal of these studies is to identify, characterize and eventually interfere with the adherence mechanisms responsible for mucosal colonization of H. influenzae type b. This application seeks to study certain surface structures (pili) of H. influenzae b that mediate adherence to human epithelial cells, and to further define the role of pili in mucosal colonization. To achieve this goal, the applicant will examine chromosomal DNA from a variety of H. influenzae type b strains to determine their homology to the structural gene of a prototypic pili of the H. influenzae b (strain M43). This gene has been cloned and sequenced by the applicant. Pili deficient (p-) mutants are being constructed by insertion of the 2 kb omega fragment encoding spectinomycin and streptomycin resistance. This mutated gene will be inserted back into the M43p+ derivative, yielding isogeneic p-transformant. The strain will be tested for its ability to adhere to epithelia and colonize respiratory mucosa in a primate model. The applicant also seeks to sequence the regions surrounding the structural gene for evidence of other genes affecting pili expression. Transposon mutagenesis of these accessory genes will be used to evaluate their role and regulation of assembly or adherence. "Such mutants will be evaluated for functional pili by electron microscopy, by adherence assays, and hemagglutination". Information obtained from these studies will clearly define the role of pili in H. influenzae b mucosal colonization. From such knowledge gained within this application, strategies can be developed to interfere with epithelial cell adherence to prevent infection.

描述：（改编自申请人摘要）。 长期目标是 这些研究是为了识别、表征并最终干扰 粘附机制负责粘膜定植H. 流感病毒B型。本申请旨在研究某些表面 结构（皮利）。流感病毒B介导对人粘附 上皮细胞，并进一步确定皮利在粘膜上皮细胞中的作用， 殖民化 为了实现这一目标，申请人将检查染色体 DNA从各种H.流感B型菌株，以确定其 同源性的原型皮利的结构基因。流感病毒B型 （菌株M43）。 该基因已由申请人克隆并测序。 通过插入2kb的peptide片段构建皮利缺陷（p-）突变体。 编码壮观霉素和链霉素抗性的Ω片段。 这 突变的基因将被插入回M43 p+衍生物中， 等基因型 该菌株将被测试其能力， 粘附于上皮细胞并在灵长类动物模型中定殖于呼吸道粘膜。 本申请人还寻求对围绕结构性寡核苷酸的区域进行测序。 基因作为影响皮利表达的其他基因的证据。 转座子 这些辅助基因的诱变将用于评估它们的作用 以及组装或粘附的调节。 “这些突变体将被评估 对于功能性皮利，通过电子显微镜、粘附试验，以及 血凝反应”。 从这些研究中获得的信息将清楚地 明确了皮利在H.流感病毒B粘膜定植。 从该等 在此应用程序中获得的知识，可以制定策略， 干扰上皮细胞粘附以防止感染。