Construction of a Vibrio cholerae vaccine candidate using transposon delivery and FLP recombinase-mediated excision
Construction of a Vibrio cholerae vaccine candidate using transposon delivery and FLP recombinase-mediated excision
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DOI：
10.1128/iai.68.11.6391-6397.2000
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发表时间：
2000-11-01
影响因子：
3.1
通讯作者：
Mekalanos, JJ
中科院分区：
文献类型：
--
作者：
Chiang, SL;Mekalanos, JJ
最近开发针对腹泻病霍乱的疫苗的努力集中在使用致病菌霍乱弧菌的减毒活菌株上。小川脂多糖表型在许多流行菌株中表达，运动缺陷降低了疫苗接种者反应性腹泻的风险。因此，我们使用携带rfbT（表达Ogawa表型所需的基因）的水手转座子，将运动Inaba（+）候选疫苗Peru-2转化为非运动Ogawa（+）表型。对22个非运动性秘鲁-2突变株的分析表明，其中2个突变株为Ogawa(+)，这2个突变株均有flgE基因插入。通过引入一种作用于转座子上抗生素标记的重组酶，有可能将这些菌株转化为抗生素敏感性。由此产生的菌株能够在幼鼠体内定植，因此可能适合作为候选疫苗单独使用或与不同生物型和血清型的菌株组合使用。
Recent efforts to develop a vaccine against the diarrheal disease cholera have focused on the use of live attenuated strains of the causative organism, Vibrio cholerae. The Ogawa lipopolysaccharide phenotype is expressed by many epidemic strains, and motility defects reduce the risk of reactive diarrhea in vaccine recipients. We therefore converted a motile Inaba(+) vaccine candidate, Peru-2, to a nonmotile Ogawa(+) phenotype using a mariner-based transposon carrying rfbT, the gene required for expression of the Ogawa phenotype. Analysis of 22 nonmotile Peru-2 mutants showed that two were Ogawa(+), and both of these strains had insertions in the flgE gene. It was possible to convert these strains to antibiotic sensitivity by introducing a recombinase that acts on sites flanking the antibiotic marker on the transposon. The resulting strains are competent for colonization in infant mice and may therefore be suitable as vaccine candidates for use either independently or in a combination with strains of different biotypes and serotypes.