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东方巴贝斯虫顶质体基因组的特征分析与注释

Characterization and annotation of Babesia orientalis apicoplast genome.

作者信息

Huang Yuan, He Lan, Hu Jinfang, He Pei, He Junwei, Yu Long, Malobi Ngabu, Zhou Yanqin, Shen Bang, Zhao JunLong

机构信息

State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, P.R. China.

Key Laboratory of Animal Epidemical Disease and Infectious Zoonoses, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.

出版信息

Parasit Vectors. 2015 Oct 16;8:543. doi: 10.1186/s13071-015-1158-x.

Abstract

BACKGROUND

Babesia orientalis is an obligate intraerythrocytic protozoan parasite of the buffalo (Bubalus bubalis, Linnaeus, 1758) transmitted by the tick Rhipicephalus heamaphysaloides. It is the causative agent of water buffalo babesiosis, one of the most important pathogens of water buffalo in central and southern China. As a member of the phylum Apicomplexa, B. orientalis possesses a relatively independent and alga originated organelle the apicoplast. Apicoplasts in other apicomplexa parasites are involved in the biosynthesis of haem, fatty acids, iron-sulphur clusters and isoprenoids. Some of these metabolic pathways were shown to be essential for parasite survival, therefore can serve as potential drug targets.

METHODS

30 pairs of primers were designed based on the full genome sequence of B. orientalis (unpublished data) and by aligning reported apicoplast genomes of Babesia bovis and Theileria parva. Conventional PCRs was performed to obtain overlapped fragments to cover the whole apicoplast genome. Then the apicoplast genome of B.orientalis was sequenced, assembled and aligned with reported apicoplast genomes of B. bovis and T. parva. The obtained apicoplast genome was annotated by using Artemis and comparing with published apicomplexan apicoplast genomes. The SSU and LSU nucleotide sequences generated were used in a phylogenetic analysis using the maximum likelihood implemented in MAGE 6.0.

RESULTS

We have obtained and analyzed the complete genome sequence of the B. orientalis apicoplast. It consisted of a 33.2 kb circular DNA (78.9 % A + T). The apicoplast genome unidirectionally encodes one large and one small subunit ribosomal RNAs, 24 tRNA genes, 4 DNA-dependent RNA polymerase beta subunits (rpoB, rpoC1, rpoC2a and rpoC2b), 17 ribosomal proteins, one EF-Tu elongation factor, 2 Clp protease chaperones, and 14 hypothetical proteins. In addition, it includes two copies of the clpC gene. The structure and organization of the B. orientalis apicoplast genome are most similar to those of the B. bovis apicoplast.

CONCLUSIONS

This is the first report of the complete sequence of the B. orientalis apicoplast genome. This information should be useful in the development of safe and efficient treatment against buffalo babesiosis.

摘要

背景

东方巴贝斯虫是水牛(Bubalus bubalis,林奈,1758)的一种专性红细胞内原生动物寄生虫,由镰形扇头蜱传播。它是水牛巴贝斯虫病的病原体,是中国中部和南部水牛最重要的病原体之一。作为顶复门的一员,东方巴贝斯虫拥有一个相对独立且起源于藻类的细胞器——顶质体。其他顶复门寄生虫的顶质体参与血红素、脂肪酸、铁硫簇和类异戊二烯的生物合成。其中一些代谢途径被证明对寄生虫的生存至关重要,因此可作为潜在的药物靶点。

方法

基于东方巴贝斯虫的全基因组序列(未发表数据),并通过比对已报道的牛巴贝斯虫和小泰勒虫的顶质体基因组,设计了30对引物。进行常规PCR以获得重叠片段,覆盖整个顶质体基因组。然后对东方巴贝斯虫的顶质体基因组进行测序、组装,并与已报道的牛巴贝斯虫和小泰勒虫的顶质体基因组进行比对。使用Artemis对获得的顶质体基因组进行注释,并与已发表的顶复门顶质体基因组进行比较。生成的SSU和LSU核苷酸序列用于在MAGE 6.0中实现的最大似然法系统发育分析。

结果

我们获得并分析了东方巴贝斯虫顶质体的完整基因组序列。它由一个33.2 kb的环状DNA组成(A+T含量为78.9%)。顶质体基因组单向编码一个大亚基和一个小亚基核糖体RNA、24个tRNA基因、4个DNA依赖性RNA聚合酶β亚基(rpoB、rpoC1、rpoC2a和rpoC2b)、17个核糖体蛋白、一个EF-Tu延伸因子、2个Clp蛋白酶伴侣和14个假定蛋白。此外,它包括两个clpC基因拷贝。东方巴贝斯虫顶质体基因组的结构和组织与牛巴贝斯虫的顶质体最为相似。

结论

这是东方巴贝斯虫顶质体基因组完整序列的首次报道。这些信息对于开发安全有效的水牛巴贝斯虫病治疗方法应该是有用 的。

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