Construction of mutants of Actinobacillus actinomycetemcomitans defective in serotype b-specific polysaccharide antigen by insertion of transposon Tn 916
Setsuko Sato1​, Nobumi Takamatsu1​, Nobuo Okahashi1​, Nobuko Matsunoshita2​, Masakazu Inoue2​, Tadamichi Takehara3​, Toshihiko Koga1​
Affiliations: 1​ Department of Dental Research, National Institute of Health, Kamiosaki, Shinagawa-ku, Tokyo 141, Japan 2​ Department of Preventive Dentistry, Kagoshima University Dental School, Sakuragaoka, Kagoshima 890, Japan 3​ Department of Preventive Dentistry, Kyushu Dental College, Kokurakitaku, Kitakyushu 803, Japan
*Author for correspondence. Tel. 03–3444–2181; fax 03–3446–6286.
First Published: 01 June 1992 https://doi.org/10.1099/00221287-138-6-1203
Mutants of Actinobacillus actinomycetemcomitans strain Y4 defective in the capsular-like serotype b-specific polysaccharide antigen (SPA) were constructed by inserting the transposon Tn 916. Southern blot analysis suggested that the transposon was inserted into a variety of different sites on the chromosome. Whole cells from two mutants (strains ST1 and ST2) lacked reactivity with a monoclonal antibody to SPA of A. actinomycetemcomitans Y4 (mAb S5) in enzyme-linked immunosorbent assay, but those from another nine mutants (e.g. strains ST3 and ST5) reacted very weakly with mAb S5. Immunodiffusion tests showed that mAb S5 or rabbit antiserum against whole cells of strain Y4 produced a fused precipitin band with purified SPA and autoclaved extract from strain Y4, but no precipitin band with autoclaved extracts from these four mutants. The hydrolysate of autoclaved extract from strain Y4 contained equal amounts of rhamnose and fucose, component sugars of SPA. The hydrolysates of autoclaved extracts from strains ST1 and ST2 contained a trace amount of rhamnose, but not fucose. Those of autoclaved extracts from strains ST3 and ST5 contained a trace amount of fucose, but not rhamnose. All of these SPA-defective mutants reacted with a mAb to lipopolysaccharide of strain Y4. The cell hydrophobicity of SPA-defective mutants was higher than that of the parent strain. These mutant clones will be useful for analysing the gene complex responsible for the synthesis of SPA of A. actinomycetemcomitans and the regulation of expression of the polysaccharide.
An erratum has been published for this content:
Construction of mutants of Actinobacillus actinomycetemcomitans defective in serotype b-specific polysaccharide antigen by insertion of transposon Tn916
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Microbiology 138, 1203 (1992); https://doi.org/10.1099/00221287-138-6-1203
10.1099/00221287-138-6-1203