Animal Health Trust, Lanwades Park, Newmarket, Suffolk, UK.
Department of Veterinary Medicine, University of Cambridge, Cambridge, UK.
Microb Genom. 2020 Apr;6(4). doi: 10.1099/mgen.0.000362. Epub 2020 Mar 31.
The availability of next-generation sequencing techniques provides an unprecedented opportunity for the assignment of gene function. subspecies is the causative agent of strangles in horses, one of the most prevalent and important diseases of equids worldwide. However, the live attenuated vaccines that are utilized to control this disease cause adverse reactions in some animals. Here, we employ transposon-directed insertion-site sequencing (TraDIS) to identify genes that are required for the fitness of in whole equine blood or in the presence of HO to model selective pressures exerted by the equine immune response during infection. We report the fitness values of 1503 and 1471 genes, representing 94.5 and 92.5 % of non-essential genes in , following incubation in whole blood and in the presence of HO, respectively. Of these genes, 36 and 15 were identified as being important to the fitness of in whole blood or HO, respectively, with 14 genes being important in both conditions. Allelic replacement mutants were generated to validate the fitness results. Our data identify genes that are important for to resist aspects of the immune response , which can be exploited for the development of safer live attenuated vaccines to prevent strangles.
下一代测序技术的出现为基因功能的确定提供了前所未有的机会。 亚种是马传染性鼻疽的病原体,是世界范围内最普遍和最重要的马属动物疾病之一。然而,用于控制这种疾病的减毒活疫苗会在某些动物中引起不良反应。在这里,我们采用转座子导向插入位点测序(TraDIS)来鉴定在整个马血中或在 HO 存在下对 的适应性至关重要的基因,以模拟感染过程中马的免疫反应施加的选择压力。我们报告了在整个血液和 HO 存在下培养后,分别有 1503 个和 1471 个基因的适应值,分别代表 亚种中非必需基因的 94.5%和 92.5%。在这两个条件下,有 14 个基因对 亚种在整个血液或 HO 中的适应性都很重要。我们生成了等位基因替换突变体来验证适应性结果。我们的数据确定了对 抵抗免疫反应某些方面至关重要的基因,这可以用于开发更安全的减毒活疫苗来预防传染性鼻疽。