PMID: 9632592Jun 25, 1998Paper

Construction and vaccine potential of acapsular mutants of Erysipelothrix rhusiopathiae: use of excision of Tn916 to inactivate a target gene

Infection and Immunity
Yoshihiro ShimojiY Yokomizo

Abstract

We previously showed that acapsular transposon Tn916 mutants of Erysipelothrix rhusiopathiae are avirulent for mice. In this study, we constructed nonreverting acapsular mutants and examined the vaccine potential of the mutants in mice. A representative acapsular transposon mutant, 33H6, was plated on selective agar containing autoclaved chlortetracycline and quinaldic acid, and two tetracycline-sensitive mutants were obtained. Sequence analysis of chromosomal regions of the mutants in which Tn916 had flanked revealed that Tn916 had spontaneously excised from the region and that the six new nucleotides, which were presumably inserted with Tn916 into 33H6 chromosome, substituted for those present at the insertion site. The mutants were confirmed to be devoid of capsular antigen by Western immunoblotting and were nonvirulent for mice (subcutaneous 50% lethal dose [LD50], >10(9) CFU). The safety and efficacy of acapsular mutants for live vaccines was further studied by using one mutant strain, named YS-1. The YS-1 bacteria were cleared from the skin sites of inoculation, livers, and spleens of the inoculated mice by 7 days after subcutaneous (s.c.) inoculation. Mice immunized s.c. with doses ranging from 2 x 10(4) to 2 x 10(8) CFU...Continue Reading

References

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Citations

Jul 5, 2012·Clinical and Vaccine Immunology : CVI·Yohsuke OgawaYoshihiro Shimoji
Mar 7, 2014·Microbiology and Molecular Biology Reviews : MMBR·Elise Darmon, David R F Leach
Mar 27, 2002·The Journal of Veterinary Medical Science·Yoshihiro ShimojiYuichi Yokomizo
Sep 23, 2000·Journal of Biotechnology·J D BoyceB Adler
Sep 8, 2009·Veterinary Microbiology·Qinning WangThomas V Riley

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