Tian Yu, Li Fangjie, Guo Jiaying, Hu Yanli, Shu Xiang, Xia Yinjun, Kang Ting, Yu Long, Liu Qin, Nie Zheng, Wang Sen, Ao Yangsiqi, An Xiaomeng, Zhao Junlong, He Lan
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, Hubei 430070, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Wuhan, Hubei 430070, China.
Parasitol Int. 2020 Aug;77:102106. doi: 10.1016/j.parint.2020.102106. Epub 2020 Mar 14.
Babesiosis caused by Babesia orientalis is one of the most serious parasitic diseases of water buffalo in the central and south part of China. Rhoptry neck proteins (RONs) are very important protein components to form a complex moving junction (MJ) which mainly participate in the invasion processes in apicomplexan parasites. Aimed to the further investigation of the function of BoRON proteins in B. orientalis, in this study, BoRON5 was characterized. A truncated 921 bp fragment of BoRON5 with predicted antigenic epitopes was cloned and inserted into pSUMO expression vector. Recombinant protein rSUMO-BoRON5 was purified from Escherichia coli. and used to produce antisera in Kunming mice. rSUMO-BoRON5 showed strong immunosignals when blotted with the positive serum from B. orientalis-infected water buffalo. Antisera raised in Kunming mice against rSUMO-BoRON5 could detect the native BoRON5 in parasite lysates. Immuofluorescence assay showed that mice antisera of rSUMO-BoRON5 could detect merozoite in B. orientalis infected water buffalo erythrocytes. This study provides useful information for the further investigation of the BoRON5 function during B. orientalis invasion of water buffalo.
由东方巴贝斯虫引起的巴贝斯虫病是中国中部和南部地区水牛最严重的寄生虫病之一。棒状体颈部蛋白（RONs）是形成复合运动连接点（MJ）的非常重要的蛋白质成分，主要参与顶复门寄生虫的入侵过程。为了进一步研究BoRON蛋白在东方巴贝斯虫中的功能，本研究对BoRON5进行了表征。克隆了具有预测抗原表位的921 bp BoRON5截短片段，并将其插入pSUMO表达载体。重组蛋白rSUMO-BoRON5从大肠杆菌中纯化出来，并用于在昆明小鼠中制备抗血清。用东方巴贝斯虫感染的水牛阳性血清印迹时，rSUMO-BoRON5显示出强免疫信号。在昆明小鼠中针对rSUMO-BoRON5产生的抗血清可以检测寄生虫裂解物中的天然BoRON5。免疫荧光分析表明，rSUMO-BoRON5的小鼠抗血清可以检测东方巴贝斯虫感染的水牛红细胞中的裂殖子。本研究为进一步研究BoRON5在东方巴贝斯虫入侵水牛过程中的功能提供了有用信息。
