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Deletion of the rfaE gene in Haemophilus parasuis SC096 strain attenuates serum resistance, adhesion and invasion

机译:副猪嗜血杆菌SC096菌株中rfaE基因的缺失减弱了血清抗药性,黏附和侵袭能力

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In Haemophilus parasuis, the lipooligosaccharide (LOS) has been identified as an important virulence factor. The rfa gene cluster encodes enzymes for LOS core biosynthesis. In order to investigate the role of the rfaE gene, we generated an rfaE deficient mutant (Delta rfaE) of a H. parasuis SC096 by a natural transformation method. The purified preparation of LOS from the Delta rfaE mutant strain showed truncated LOS structure on silver-stained SOS-PAGE. Compared to the wild-type SC096 strain, the generation time of Delta rfaE mutant strain was significantly extended from 59 min to 69 min. The Delta rfaE mutant strain caused an approximately 30-fold reductions in survival rate in 50% sera and 36-fold reductions in survival rate in 90% sera, respectively (p 0.001). In adhesion and invasion assays, the Delta rfaE mutant strain had 10-fold less efficient adherence and 12-fold reductions in invasion of the porcine umbilicus vein endothelial cells (PUVEC) and porcine kidney epithelial cells (PK-15), respectively (p 0.001). However, the complemented strain could restore the above phenotypes. Hence, the above results suggested that the rfaE gene participated in the pathogenicity of H. parasuis SC096 strain. (C) 2014 Elsevier Ltd. All rights reserved.
机译:在副猪嗜血杆菌中,脂寡糖(LOS)被确定为重要的毒力因子。 rfa基因簇编码用于LOS核心生物合成的酶。为了研究rfaE基因的作用,我们通过自然转化方法生成了副猪嗜血杆菌SC096的rfaE缺陷突变体(Delta rfaE)。从Delta rfaE突变株纯化的LOS制备物在银染的SOS-PAGE上显示出截短的LOS结构。与野生型SC096菌株相比,Delta rfaE突变株的产生时间从59分钟显着延长至69分钟。 Delta rfaE突变株分别导致50%血清存活率降低约30倍和90%血清存活率降低36倍(p <0.001)。在粘附和侵袭试验中,Delta rfaE突变株对猪脐静脉血管内皮细胞(PUVEC)和猪肾上皮细胞(PK-15)的粘附效率分别降低10倍和12倍(p < 0.001)。但是,互补菌株可以恢复上述表型。因此,以上结果提示rfaE基因参与了副猪嗜血杆菌SC096株的致病性。 (C)2014 Elsevier Ltd.保留所有权利。

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