...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Infection and immunity >Construction and characterization of a potential live oral carrier-based vaccine against Vibrio cholerae O139.
【24h】

Construction and characterization of a potential live oral carrier-based vaccine against Vibrio cholerae O139.

机译：抗霍乱弧菌O139的潜在的基于口服载体的活疫苗的构建和表征。

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The rfb region from Vibrio cholerae O139 strain MO45 was cloned from cosmid gene banks established in Escherichia coli HB101, using an immunoblot assay for screening of the correct clones. Immunoblot analysis of lipopolysaccharide (LPS) preparations revealed the presence of two types of positive clones: (i) those expressing only a short core-linked O polysaccharide (SOPS) and (ii) those also expressing a highly polymerized capsular polysaccharide (CPS) not bound to the E. coli K-12 LPS core. In addition, the latter clones appear to contain a locus which may encode a putative regulator of SOPS and CPS chain length. Further characterization in E. coli showed that CPS constitutes a barrier against large particles such as the bacteriophage Ffm but not against bacteriophage lambda or P1. In addition, a portion of the K-12 LPS core may not be substituted with SOPS. Loci associated with the two clonal types were transferred into V. cholerae CH19, an rfbAB deletion mutant of CVD103-HgR deficient in the production of the homologous Inaba O polysaccharide. This resulted in the stable expression of SOPS, alone or together with CPS, that was indistinguishable from that of wild-type V. cholerae O139. Strains CH25 and CH26, which correspond to CH19 bearing the V. cholerae O139 rfb region integrated into the chromosome, were found to be genetically stable and essentially identical to the parent CVD103-HgR with respect to physiological properties such as cell motility, mercury resistance, toxicity, and production of the cholera toxin B subunit. Rabbits immunized with CH25 elicited high titers of anti-O139 SOPS- and CPS-specific serum antibodies. These strains possess characteristics desirable in candidate live oral vaccines against V. cholerae O139.
机译：使用免疫印迹试验筛选正确的克隆，从霍乱弧菌O139菌株MO45的rfb区克隆自在大肠杆菌HB101中建立的粘粒基因库。脂多糖（LPS）制剂的免疫印迹分析表明存在两种类型的阳性克隆：（i）仅表达短核心连接的O多糖（SOPS）的克隆，以及（ii）也表达高度聚合的荚膜多糖（CPS）的克隆绑定到大肠杆菌K-12 LPS核心。另外，后面的克隆似乎含有一个基因座，该基因座可以编码一个假定的SOPS和CPS链长调节子。在大肠杆菌中的进一步表征表明，CPS构成了对大颗粒（如噬菌体Ffm）的屏障，但对λ噬菌体或P1则不是屏障。另外，K-12 LPS核心的一部分可能不会被SOPS取代。与这两种克隆类型相关的基因座被转移到霍乱弧菌CH19中，霍乱弧菌CH19是缺乏同源Inaba O多糖生产的CVD103-HgR的rfbAB缺失突变体。这导致SOPS单独或与CPS一起稳定表达，这与野生型霍乱弧菌O139的表达没有区别。发现对应于带有整合入染色体的霍乱弧菌O139 rfb区的CH19的菌株CH25和CH26具有遗传稳定性，并且在生理特性（例如细胞运动性，耐汞性，毒性和霍乱毒素B亚基的产生。用CH25免疫的兔子引起高滴度的抗O139 SOPS和CPS特异性血清抗体。这些菌株具有抗霍乱弧菌O139的候选口服活疫苗中所需的特性。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号