Identification of a genetic locus essential for serotype b-specific antigen synthesis in Actinobacillus actinomycetemcomitans

Yasuo Yoshida, Yoshio Nakano, Yoshihisa Yamashita, Toshihiko Koga

研究成果: Contribution to journalArticle査読

43 被引用数 (Scopus)

抄録

A large gene cluster associated with the biosynthesis of the serotype- specific polysaccharide antigen (SPA) of Actinobacillus actinomycetemcomitans Y4 (serotype b) was cloned and characterized. Western blot analysis showed that Escherichia coli DHSα, containing a plasmid carrying this cluster, produced a polysaccharide which reacted with a monoclonal antibody directed against the SPA of A. actinomycetemcomitans Y4. High-performance liquid chromatography analysis indicated that the polysaccharide produced by an E. coli transformant, as well as A. actinomycetemcomitans Y4 SPA, was composed of rhamnose and fucose. Furthermore, using various derivatives of the plasmid, we demonstrated that the cloned 13-kb BssHII-BspHI fragment was indispensable for SPA synthesis in E. coli DH5α. The 24,909-bp nucleotide sequence, which included this fragment and its flanking regions, was determined. In the sequenced area, 24 open reading frames (ORFs) with the same orientation were found. Most of these were located sequentially within a short distance of each other. Many of the deduced amino acid sequences were similar to the gene products of the polysaccharide synthetic genes of other bacteria. The average G+C content (37.7%) of all 24 ORFs in the sequenced area was lower than that (45.6%) of the whole chromosome of A. actinomycetemcomitans Y4. It is noteworthy the average G+C content of the nine ORFs in the 8.5-kb central region of the 13-kb BssHII-BspHI fragment indispensable for SPA synthesis in E. coli was found to be especially low (27.0%).

本文言語英語
ページ(範囲)107-114
ページ数8
ジャーナルInfection and Immunity
66
1
DOI
出版ステータス出版済み - 1998

All Science Journal Classification (ASJC) codes

  • 寄生虫科
  • 微生物学
  • 免疫学
  • 感染症

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