Abstract：
In humans, Entamoeba histolytica is the main pathogen causing various amoebiases, while E. moshkovskii falls between being a pathogen and non-pathogen. The two species have similar behavior patterns but differ significantly in pathogenicity, with previous studies and clinical data indicating that E. moshkovskii has a low level of pathogenicity. Meaningfully, the biological characteristics of E. moshkovskii make it a potential model organism and a protein display platform for studying the functions of important Entamoeba proteins. Here, an Amoeba-pcDNA3.1 vector capable of overexpressing E. histolytica-sourced Igl-C protein was constructed and successfully transfected into E. moshkovskii. High levels of expression of the Igl-C, EGFP, and NeoR genes were identified in Igl-C-transfected trophozoites using qRT-PCR, and they were subsequently confirmed using immunoblotting. Transfection of Igl-C protein improved the adherence and phagocytosis of E. moshkovskii, demonstrating that E. histolytica Igl mediated amoebic adhesion. Moreover, as a manifestation of protein virulence, the ability of post-transfected trophozoites to induce inflammation in host macrophages was also enhanced. In conclusion, this study utilizing the characteristics of E. moshkovskii confirmed its potential to serve as a model organism. E. moshkovskii could replace E. histolytica as the target of gene editing, allowing more efficient study of amoebic pathogenicity.
摘要：
在人类中,溶组织内阿米巴是引起各种变形虫的主要病原体，而E.moshkovskii介于病原体和非病原体之间。这两个物种具有相似的行为模式，但在致病性方面存在显着差异。先前的研究和临床数据表明莫什科夫斯基大肠杆菌具有低水平的致病性。有意义的,E.moshkovskii的生物学特性使其成为潜在的模型生物和蛋白质展示平台，用于研究重要的Entamoeba蛋白的功能。这里,构建能够过表达溶组织性大肠杆菌来源的Igl-C蛋白的变形虫-pcDNA3.1载体,并成功转染入E.moshkovskii。Igl-C的高水平表达，EGFP,使用qRT-PCR在Igl-C转染的滋养体中鉴定NeoR基因，随后使用免疫印迹进行了确认。Igl-C蛋白的转染改善了E.moshkovskii的粘附和吞噬作用，证明溶组织大肠杆菌Igl介导阿米巴粘附。此外，作为蛋白质毒力的表现，转染后滋养体诱导宿主巨噬细胞炎症的能力也增强.总之,这项研究利用了E.moshkovskii的特征，证实了其作为模型生物的潜力。E.moshkovskii可以取代溶组织大肠杆菌作为基因编辑的靶标，允许更有效地研究阿米巴致病性。
