Correlation between infection mechanisms and virulence plasmid of Salmonella choleraesuis

猪霍乱沙门氏菌感染机制与毒力质粒的相关性

基本信息

批准号：
63570205
负责人：
DANBARA Hirofumi
金额：
$ 1.34万
依托单位：
The Kitasato Institute
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63570205/
关键词：
Salmonella choleraesuis Salmonella Virulence plasmid Bacteremia 毒力プラスミド

项目摘要

1. Correlation between virulence plasmid of Salmonella choleraesuis, pKDSC50, and its mouse virulence: S. choleraesuis strains harboring virulence plasmid, pKDSC50 (parent and re-introduced strains), and cured strains were inoculated intraperitoneally into ICR mice, and the mice were examined pathologically for 14 days. Parent and re-introduced strains caused bacteremia in mice, and inoculated bacteria were recovered from spleen and liver. Obvious pathological changes were observed at individual tissues of peritoneal fluid, mesenterium, and visceral organs, and those changes persisted to day 14. In contrast, mice infected with plasmid-cured strain showed no bacteremia, and had only mild and transient pathological changes at peritoneal fluid, mesenterium, and viscera.2. Difference of cell surface components between pKDSC50-harbouring and -cured strains of S. choleraesuis : Lipopolysaccharide, outer membrane proteins, and phospholipids of parent, re-introduced, and cured strains were che … More mically and biochemically compared. There was no significant difference between them, indicating that pKDSC50 did not mediate the detectable change of bacterial cell surface.3. Mapping of the region responsible for the ability to cause mouse bacteremia by transposoninsertional inactivation: Mouse bacteremia test was established in the base of the results of pathological study, and used for screening 100 mutant strains constructed by transposon Tnl-insertional inactivation. Plasmid DNA of mouse bacteremia test-negative strains were introduced to cured strain by transformation, and the transformants were again tested for mouse bacteremia ability. Nine strains were negative for the second test, and Tnl on their plasmids were localized in 6.2 kb region of pKDSC50. This region was designated mba region.4. Genetical study on mba region of pKDSC50: DNA fragments of mba region were cloned in E. coli, and proteins encoded in this region were identified by E. coli minicell system, or by lac-promoter system with induction of isopropylthiogalactoside. Four proteins with apparent molecular weights of 29k, 70k, 32k, and 32k were detected, and open reading frames corresponding to these four proteins were found in mba region by nucleotide sequential analysis. Other research groups has recently reported similar proteins encoded in virulence plasmid of S. typhimurium, suggesting that the proteins we found were common among virulence plasmid-harboring Salmonella, and playing important role in pathogenesis of Salmonella. Less

1。沙门氏菌霍乱的病毒质粒，PKDSC50及其小鼠病毒：具有病毒质粒，PKDSC50（父母和重新引起的菌株）的霍乱链球菌菌株，并在ICR小鼠内进行了腹腔内的接种，并在ICR小鼠内接种了治疗菌株，并进行了固定的菌株，并进行了14天的病理学。父母和重新引入的菌株在小鼠中引起细菌，并从脾脏和肝脏中回收了接种的细菌。在腹膜液，肠系膜和内脏器官的单个组织中观察到了明显的病理变化，这些变化持续到第14天。相反，被质粒蛋白菌株感染的小鼠没有细菌，并且仅在腹膜液体，细胞植物，伴有腹膜液体的病理学变化中，并且只有轻度的病理学变化。细胞表面成分之间的细胞表面成分与霍乱链球菌的PKDSC50-HARBORING菌株：脂多糖，外膜外蛋白以及父母的磷脂，重新引入和固化的菌株是CHE…更模拟和生物学上比较。它们之间没有显着差异，表明PKDSC50并未介导细菌细胞表面的可检测到的变化3。在病理学研究结果的基础上建立了负责通过转钉灭活引起小鼠细菌的区域的映射：小鼠细菌测试，并用于筛选由转孔TNL-TNL插入式失活构建的100个突变菌株。小鼠细菌测试阴性菌株的质粒DNA通过转化引入固化菌株，并再次测试了转化因子的小鼠细菌能力。在第二次测试中，有9个菌株为阴性，其质粒上的TNL位于PKDSC50的6.2 kb区域中。该地区被指定为MBA地区4。对PKDSC50的MBA区域的遗传研究：将MBA区域的DNA片段克隆在大肠杆菌中，并通过大肠杆菌微型细胞系统或通过诱导异丙基硫代糖苷的诱导，在该区域编码了该区域的蛋白质。通过核苷酸顺序分析发现了四种具有29K，70K，32K和32K分子量的蛋白质，并在MBA区域发现了与这四个蛋白相对应的开放式阅读框。其他研究组最近报道了在伤寒链霉菌病毒质粒中编码的类似蛋白质，这表明我们发现的蛋白质在病毒质粒 - 荷兰沙门氏菌中很常见，并且在沙门氏菌的发病机理中起重要作用。较少的