Ma Lei, Liu Jing, Li Muzi, Fu Yong, Zhang Xiao, Liu Qun
Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University Beijing, China.
Front Microbiol. 2017 Mar 7;8:370. doi: 10.3389/fmicb.2017.00370. eCollection 2017.
, of the phylum, is a common cause of abortions in cattle and nervous system dysfunction in dogs. Rhoptry proteins of play an important role in virulence. The objectives of this study were to study functions of NcROP5 in by deleting the NcROP5 gene from the wild Nc-1 strain. We selected in ToxoDB and successfully constructed an gene-deleted vector, pTCR-NcROP5-CD KO. Then we screened the NcROP5 knockout strains (ΔNcROP5) at the gene, protein and transcription levels. Plaque assay, host cell invasion assay and intracellular proliferation test showed that the ΔNcROP5 strain had less plaque space, weakened invasion capacity and slower intracellular growth. Animal testing showed significantly lower cerebral load of ΔNcROP5 than the load of the Nc-1 strain, as well as a loss of virulence for the ΔNcROP5 strains. Phenotypic analyses using the label-free LC-MS/MS assay-based proteomic method and KEGG pathway enrichment analysis showed a reduction of transcription and altered expression of multiple proteins including the apicomplexan family of binding proteins. The present study indicated that ROP5 is a key virulence factor in in mice. The proteomic profiling of Nc-1 and ΔNcROP5 provided some data on differential proteins. These data provide a foundation for future research of protein functions in .
属于该门的(寄生虫名称未给出)是牛流产和犬神经系统功能障碍的常见病因。(寄生虫名称未给出)的棒状体蛋白在毒力方面发挥重要作用。本研究的目的是通过从野生Nc - 1株中删除NcROP5基因来研究其在(寄生虫名称未给出)中的功能。我们在ToxoDB中进行筛选并成功构建了一个NcROP5基因缺失载体,即pTCR - NcROP5 - CD KO。然后我们在基因、蛋白质和转录水平筛选了NcROP5基因敲除株(ΔNcROP5)。噬斑测定、宿主细胞侵袭测定和细胞内增殖试验表明,ΔNcROP5株的噬斑面积较小、侵袭能力减弱且细胞内生长较慢。动物试验表明,ΔNcROP5的脑内负荷明显低于Nc - 1株,并且ΔNcROP5株丧失了毒力。使用基于无标记LC - MS/MS分析的蛋白质组学方法和KEGG通路富集分析进行的表型分析表明,(寄生虫名称未给出)转录减少以及包括顶复门结合蛋白家族在内的多种蛋白质表达改变。本研究表明,ROP5是(寄生虫名称未给出)在小鼠中的关键毒力因子。Nc - 1和ΔNcROP5的蛋白质组图谱提供了一些差异蛋白的数据。这些数据为未来对(寄生虫名称未给出)中蛋白质功能的研究奠定了基础。