LIPOOLIGOSACCHARIDE BIOSYNTHESIS IN NEISSERIACEAE

奈瑟菌科中脂寡糖的生物合成

• 批准号: 2062587
• 负责人: DANIEL C STEIN
• 金额: $ 18.01万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-30 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2062587
• 关键词: Escherichia coli; SDS polyacrylamide gel electrophoresis; bacterial DNA; bacterial antigens; bacterial genetics; carbohydrate biosynthesis; drug resistance; endotoxins; galactose; gene complementation; gene deletion mutation; genetic regulation; genetic strain; lipopolysaccharides; microorganism metabolism; molecular cloning; mutant; oligosaccharides; plasmids; protein purification; recombinant DNA; sialyltransferases; transposon /insertion element

Neisseria gonorrhoeae is responsible for over l million cases of gonorrhea each year, and the total health care costs associated with treating gonorrhea and complications that arise from infections caused by this organism exceed l billion dollars per year. A closely related pathogen, N. meningitidis, causes significant morbidity and mortality worldwide. If effective vaccines to prevent these diseases were available, health care costs would be dramatically reduced. The gonococcus possesses a wide variety of surface structures that can serve as virulence determinants for pathogenesis. Lipooligosaccharide (LOS) is the least understood cell surface antigen, both chemically and genetically, even though these molecules mediate most of the toxic damage in fallopian tubes, are responsible for complement activation on the cell surface, and serve as the target for bactericidal antibody. Furthermore, antibody directed against lipopolysaccharide is protective for a variety of Gram-negative infections. A vaccine that stimulated an anti-LOS response might be protective for both gonococcal and meningococcal infections. Since gonococcal and meningococcal LOS share many epitopes, and all of the genes that have been identified that are involved in LOS biosynthesis are conserved in both species, information gleaned about LOS biosynthesis in one species will be directly applicable to the other. This proposal will focus on LOS biosynthesis in the gonococcus. It will identify by gene cloning and genetic complementation techniques, genes required for LOS biosynthesis, characterize the genes by DNA sequence analysis, define the biochemical properties of the gene products and determine the mechanisms by which the gonococcus regulates their synthesis. If LOS is to be exploited as a vaccine candidate, we need to have strains of the gonococcus that stably produce the desired epitopes. Since the gonococcus can vary its LOS, depending on the host, and local environment, studies on the genetic regulation of its synthesis are warranted. Well defined, stable LOS producing strains are needed if we are to dissect the role of LOS in the disease process. By understanding how the gonococcus makes LOS, it will be possible to construct strains with defined LOS. These strains will be the basis of future studies into the role of LOS in the disease process.

淋病奈瑟菌导致超过一百万例 淋病每年，以及与之相关的总医疗费用 治疗淋病和由感染引起的并发症， 每年超过10亿美元。一个密切相关 pathogenic、N.脑膜炎，造成严重的发病率和死亡率 国际吧如果有效的疫苗能预防这些疾病， 医疗保健费用将大大降低。的 淋球菌具有多种多样的表面结构， 作为致病的毒力决定因素。低聚脂（LOS）是 最不了解的细胞表面抗原，化学上和 从遗传学上讲，尽管这些分子介导了大部分的毒性损伤， 在输卵管中，负责细胞上的补体激活 表面，并作为杀菌抗体的目标。此外，委员会认为， 针对脂多糖的抗体对多种 革兰氏阴性菌感染一种能激发抗LOS抗体的疫苗 对淋球菌和脑膜炎球菌的免疫应答可能具有保护作用 感染.由于淋球菌和脑膜炎球菌LOS共享许多表位， 所有已经被确认的与LOS相关的基因 生物合成在两个物种中都是保守的，收集到的有关LOS的信息 一个物种的生物合成将直接适用于另一个物种。 该建议将集中于淋球菌中LOS的生物合成。它将 通过基因克隆和基因互补技术鉴定， LOS生物合成所必需的基因，通过DNA序列表征基因 分析，确定基因产物的生化特性， 确定淋球菌调节其 合成.如果LOS被开发为候选疫苗，我们需要 具有稳定产生所需表位的淋球菌菌株。 由于淋球菌可以改变其LOS，这取决于宿主和局部 环境，研究其合成的遗传调控是 有正当理由如果我们需要良好定义的，稳定的LOS生产菌株， 是剖析LOS在疾病过程中的作用。通过了解 淋球菌是如何制造LOS的， 定义的LOS。这些菌株将是未来研究的基础， LOS在疾病过程中的作用。