Identification of a novel thrombospondin-related anonymous protein (BoTRAP2) from Babesia orientalis
Identification of a novel thrombospondin-related anonymous protein (BoTRAP2) from Babesia orientalis
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东方巴贝虫中新型血小板反应蛋白相关匿名蛋白 (BoTRAP2) 的鉴定
DOI：
10.1186/s13071-019-3457-0
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发表时间：
2019-05
影响因子：
3.2
通讯作者：
Zhao Junlong
中科院分区：
文献类型：
--
作者：
Zhan Xueyan;He Junwei;Yu Long;Liu Qin;Sun Yali;Nie Zheng;Guo Jiaying;Zhao Yangnan;Li Muxiao;Luo Xiaoying;He Lan;Zhao Junlong
背景：凝血酶原蛋白相关匿名蛋白(TRAP)最先在恶性疟原虫的子孢子中发现，TRAP家族蛋白由微线体分泌并转运到寄生虫表面参与入侵过程。在尖端复合体原生动物中已鉴定出多种TRAP蛋白，但有关东方巴贝斯虫TRAP2蛋白的报道较少。方法：克隆东方巴贝斯虫TRAP2的功能结构域，并与相关尖端复合体寄生虫的TRAP序列进行比较。将BoTRAP2的功能结构域截短，命名为BoTRAP2-1，并将其克隆到表达载体pET-28a中。免疫3只兔，制备兔抗rBoTRAP2-1多克隆抗体。Western印迹分析鉴定BoTRAP2的天然形式和免疫原性。结果：扩增的BoTRAP2基因全长2817个碱基，编码一个约938个氨基酸的功能结构域，包括两个VWFA结构域，一个是TSP结构域，一个是跨膜结构域。BoTRAP2的氨基酸序列与牛黄杆菌和吉氏杆菌有很高的相似性。预测的截短的BoTRAP2-1的三级结构证实了BoTRAP2含有两个VWFA结构域和一个TSP结构域，这是该蛋白质的主要功能区域。用兔多克隆抗体rBoTRAP2-1从侧柏裂解液中鉴定出天然的BoTRAP2。与水牛感染水牛血清反应，检测到一条与rBoTRAP2-1相对应的条带，表明该蛋白具有较高的免疫原性。免疫荧光分析表明，兔抗rBoTRAP2-1多克隆血清可将BoTRAP2主要定位于寄生虫的顶端。结论：该rBoTRAP2能区分感染水牛和正常水牛的血清，提示BoTRAP2具有较高的免疫原性，可作为诊断水牛东方杆菌感染的候选抗原。
Background:The thrombospondin-related anonymous protein (TRAP) was first discovered in the sporozoite of Plasmodium falciparum and TRAP family proteins are secreted by micronemes and transported to the parasite surface to participate in the invasion process. Various TRAP proteins have been identified in apicomplexan protozoans, but there have been few reports about TRAP proteins in Babesia orientalis.Methods:The functional domain of TRAP2 in B. orientalis was cloned, sequenced, characterized and compared to the TRAP sequences of related apicomplexan parasites. The functional domain of BoTRAP2 was truncated, named BoTRAP2-1, and then cloned into the pET-28a expression vector. Rabbit anti-rBoTRAP2-1 polyclonal antibody was produced by immunizing three rabbits. Western blot analysis was used to identify the native form and immunogenicity of BoTRAP2. The localization of BoTRAP2 was identified by indirect fluorescence assay (IFA).Results:The amplified genes of BoTRAP2 are 2817 bp in length, encoding a functional domain of about 938 aa with two vWFA domains, one TSP domain and one transmembrane domain. The amino acid sequence of BoTRAP2 has a high similarity with that of B. bovis and B. gibsoni. The predicted tertiary structure of truncated BoTRAP2-1 confirmed that BoTRAP2 contains two vWFA domains and a TSP domain, the main functional areas of the protein. The native BoTRAP2 was identified from B. orientalis lysate by using rabbit polyclonal anti-rBoTRAP2-1. A band corresponding to rBoTRAP2-1 was detected by reaction with serum from a B. orientalis-infected water buffalo, indicating that the protein has a high immunogenicity. IFA showed that BoTRAP2 is mainly localized on the apical end of parasites by rabbit anti-rBoTRAP2-1 polyclonal serum.Conclusions:The rBoTRAP2 could differentiate serum from B. orientalis-infected water buffalo and normal water buffalo, implicating that BoTRAP2 has high immunogenicity and could serve as a candidate antigen for diagnosis of B. orientalis infection in buffalo.
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影响因子：
1.7
作者：
Zhongling Liu;Junlong Zhao;Lihua Ma;B. Yao
通讯作者：
Zhongling Liu;Junlong Zhao;Lihua Ma;B. Yao
影响因子：
6.7
作者：
Kehrer J;Singer M;Lemgruber L;Silva PA;Frischknecht F;Mair GR
通讯作者：
Mair GR
影响因子：
1.5
作者：
Lovett, JL;Howe, DK;Sibley, LD
通讯作者：
Sibley, LD
影响因子：
4.8
作者：
Baum, J;Richard, D;Cowman, AF
通讯作者：
Cowman, AF
影响因子：
3
作者：
Akhouri RR;Sharma A;Malhotra P;Sharma A
通讯作者：
Sharma A