Abstract

Toxoplasmosis is a widespread zoonosis with an impact on immunocompromised people and critical in pregnant women because of its transmission to the fetus. Recombinant Toxoplasma gondii antigens produced in Escherichia coli are useful for antibody detection in patient’s blood. The aim of the study was to evaluate the feasibility of deriving chromosome-edited E. coli clones producing T. gondii antigens. Here, we present the CRISPR-Cas9 facilitated editing of the E. coli genome to produce SAG2 and GRA2 T. gondii antigens. Moreover, we have derived a clone that produces both proteins and an additional clone producing a novel fusion protein, SAG2-GRA2. These proteins, bearing His-tag, can be easily purified and are useful to detect anti-Toxoplasma antibodies in human blood. We conclude that it is feasible to edit the E. coli chromosome to produce T. gondii antigens that are bound by antibodies present in people affected by toxoplasmosis.

中文翻译：

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基因组编辑大肠杆菌中弓形虫重组抗原的生产

摘要

弓形虫病是一种广泛的人畜共患病，对免疫功能低下的人有影响，并且由于其传播给胎儿而对孕妇至关重要。在大肠杆菌中产生的重组弓形虫抗原可用于检测患者血液中的抗体。该研究的目的是评估获得产生刚地弓形虫抗原的染色体编辑的大肠杆菌克隆的可行性。在这里，我们介绍了CRISPR-Cas9方便的大肠杆菌基因组编辑以产生SAG2和GRA2 T. gondii抗原。此外，我们已经得到了一个既生产蛋白质的克隆，又一个生产新型融合蛋白SAG2-GRA2的克隆。这些带有His标签的蛋白质可以轻松纯化，可用于检测人血中的抗弓形虫抗体。我们得出结论，编辑大肠杆菌染色体以产生弓形虫抗原是可行的，该抗原与弓形虫病患者体内存在的抗体结合。