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

• 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

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.猪霍乱沙门氏菌毒力质粒pKDSC 50与小鼠毒力的相关性研究将携带毒力质粒pKDSC 50的猪霍乱菌株（亲本菌株和再引入菌株）和治愈菌株腹腔接种于ICR小鼠，并对小鼠进行病理检查14天。亲本菌株和重新引入的菌株引起小鼠菌血症，从脾脏和肝脏中回收接种的细菌。在腹膜液、肠系膜和内脏器官的单个组织中观察到明显的病理变化，并且这些变化持续至第14天。而质粒治愈株感染的小鼠则无菌血症，仅在腹腔液、肠系膜和内脏出现轻度和一过性的病理变化.携带pKDSC 50的菌株与未治愈的菌株细胞表面成分的差异。猪霍乱：检测了亲本、再引入和治愈菌株的脂多糖、外膜蛋白和磷脂， ...更多信息 从化学和生物化学上比较。两者之间无显著性差异，表明pKDSC 50不介导细菌细胞表面可检测的变化.转座子插入失活引起小鼠菌血症能力区域的定位：根据病理学研究结果建立小鼠菌血症试验，筛选100株转座子Tnl插入失活突变株。将小鼠菌血症试验阴性菌株的质粒DNA通过转化法导入治愈菌株，并对转化子进行小鼠菌血症能力的再次检测。第二次检测9株菌均为阴性，质粒上TnI定位于pKDSC 50的6.2kb区域。这个地区被指定为mba地区。pKDSC 50 mba区的遗传学研究：将mba区的DNA片段克隆到E. coli中表达，并通过E. coli微细胞系统，或通过具有异丙基硫代半乳糖苷诱导的lac-promoter系统。检测到表观分子量分别为29 k、70 k、32 k和32 k的4种蛋白质，核苷酸序列分析在mba区发现了与这4种蛋白质相对应的开放阅读框。其他研究小组最近也报道了在沙门氏菌毒力质粒中编码的类似蛋白。结果表明，这些蛋白质在携带毒力质粒的沙门氏菌中普遍存在，在沙门氏菌的致病过程中起重要作用。少

项目成果

K.Kawahara,T.Hamaoka,S.Suzuki,M.Nakamura,S.Y.Murayama,T.Arai,N.Terakado,H.Danbara: "Lipopolysaccharide alteration mediated by the virulence plasmid of Salmonella" Microbial Pathogenesis. 7. 195-202 (1989)

K.Kawahara，T.Hamaoka，S.Suzuki，M.Nakamura，S.Y.Murayama，T.Arai，N.Terakado，H.Danbara：“沙门氏菌毒力质粒介导的脂多糖改变”微生物发病机制。

H. Matsui, K. Kawahara, N. Terakado, H. Danbara.: "Nucleotide sequence of genes encoding 32 kDa and 70 kDa polypeptides in mba region of the virulence plasmid, pKDSC50, of Salmonella choleraesuis." Nucleic Acids Research.

H. Matsui、K. Kawahara、N. Terakado、H. Danbara.：“猪霍乱沙门氏菌毒力质粒 pKDSC50 的 mba 区域中编码 32 kDa 和 70 kDa 多肽的基因的核苷酸序列。”

R.Moriguchi,K.Kawahara,Y.Haraguchi,Y.Nakase,H.Danbara: "Pathological study on virulence of Salmonella choleraesuis in mice associated with 50-kilobase plasmid" British J.Experimental Pathology.

R.Moriguchi、K.Kawahara、Y.Haraguchi、Y.Nakase、H.Danbara：“与 50 kilobase 质粒相关的小鼠霍乱沙门氏菌毒力的病理学研究”英国 J.实验病理学。

R. Moriguchi, K. Kawahara, Y. Haraguchi, Y. Nakase, H. Danbara.: "Pathological study on virulence of Salmonella choleraesuis in mice associated with 50-kilobase plasmid." British J. Experimental Pathology.

R. Moriguchi、K. Kawahara、Y. Haraguchi、Y. Nakase、H. Danbara.：“与 50-kilobase 质粒相关的小鼠霍乱沙门氏菌毒力的病理学研究。”

H.Matsui,K.Kawahara,N.Terakado,H.Danbara: "Nucleotide sequence of a gene encoding a 29 KDa polypeptide in mba region of the virulence plasmid,pKDSC50,of Salmonella choleraesuis" Nucleic Acids Research. 18. (1990)

H.Matsui、K.Kawahara、N.Terakado、H.Danbara：“猪霍乱沙门氏菌毒力质粒 pKDSC50 的 mba 区域中编码 29 KDa 多肽的基因的核苷酸序列”核酸研究。