Sidhu-Muñoz Rebeca S, Sancho Pilar, Vizcaíno Nieves
Departamento de Microbiología y Genética, Edificio Departamental, Universidad de Salamanca, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain; Instituto de Investigación Biomédica de Salamanca (IBSAL), 37007 Salamanca, Spain.
Departamento de Microbiología y Genética, Edificio Departamental, Universidad de Salamanca, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain.
Vet Microbiol. 2016 Apr 15;186:59-66. doi: 10.1016/j.vetmic.2016.02.010. Epub 2016 Feb 20.
Mutants in several genes have been obtained on the genetic background of virulent rough (lacking O-polysaccharide) Brucella ovis PA. The target genes encode outer membrane proteins previously associated with the virulence of smooth (bearing O-polysaccharide chains in the lipopolysaccharide) Brucella strains. Multiple attempts to delete omp16, coding for a homologue to peptidoglycan-associated lipoproteins, were unsuccessful, which suggests that Omp16 is probably essential for in vitro survival of B. ovis PA. Single deletion of omp10 or omp19-that encode two other outer membrane lipoproteins--was achieved, but the simultaneous removal of both genes failed, suggesting an essential complementary function between both proteins. Two other deletion mutants, defective in the Tol-C-homologue BepC or in the SP41 adhesin, were also obtained. Surprisingly when compared to previous results obtained with smooth Brucella, none of the B. ovis mutants showed attenuation in the virulence, either in the mouse model or in cellular models of professional and non-professional phagocytes. Additionally, and in contrast to the observations reported with smooth Brucella strains, several properties related to the outer membrane remained almost unaltered. These results evidence new distinctive traits between naturally rough B. ovis and smooth brucellae.
在强毒株粗糙型(缺乏O-多糖)绵羊布鲁氏菌PA的遗传背景下,已获得多个基因的突变体。这些靶基因编码的外膜蛋白先前与光滑型(脂多糖中带有O-多糖链)布鲁氏菌菌株的毒力相关。多次尝试删除编码肽聚糖相关脂蛋白同源物的omp16均未成功,这表明Omp16可能对绵羊布鲁氏菌PA的体外存活至关重要。成功实现了编码另外两种外膜脂蛋白的omp10或omp19的单基因缺失,但同时删除这两个基因却失败了,这表明这两种蛋白之间存在必需的互补功能。还获得了另外两个缺失突变体,它们分别在Tol-C同源物BepC或SP41黏附素中存在缺陷。令人惊讶的是,与先前用光滑型布鲁氏菌获得的结果相比,无论是在小鼠模型还是在专业和非专业吞噬细胞的细胞模型中,绵羊布鲁氏菌的突变体均未表现出毒力减弱。此外,与光滑型布鲁氏菌菌株的观察结果相反,一些与外膜相关的特性几乎没有改变。这些结果证明了天然粗糙型绵羊布鲁氏菌与光滑型布鲁氏菌之间新的独特特征。