我国羊巴贝斯虫trap基因结构及遗传进化分析

doi:10.11843/j.issn.0366-6964.2017.07.018

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (7): 1332-1341.doi: 10.11843/j.issn.0366-6964.2017.07.018

我国羊巴贝斯虫trap基因结构及遗传进化分析

何欣, 刘军龙, 刘爱红, 王锦明, 牛庆丽, 李有全, 殷宏, 罗建勋*, 关贵全*

中国农业科学院兰州兽医研究所/家畜疫病病原生物学国家重点实验室/ 甘肃省动物寄生虫病重点实验室, 兰州 730046

收稿日期:2017-02-10 出版日期:2017-07-23 发布日期:2017-07-23
通讯作者: 关贵全,研究员,E-mail:guanguiquan@caas.cn;罗建勋,研究员,E-mail:luojianxun@caas.cn
作者简介:何欣(1992-),女,四川德阳人,硕士生,主要从事动物寄生虫及其分子生物学研究,E-mail:hexinwyyx@126.com
基金资助:
国家重点基础研究发展计划（973）项目（2015CB150300）；国家自然科学基金（31072130）；农业科技创新工程（ASTIP）

The Structural and Phylogenetic Analysis of trap Gene in Ovine Babesia Species in China

HE Xin, LIU Jun-long, LIU Ai-hong, WANG Jin-ming, NIU Qing-li, LI You-quan, YIN Hong, LUO Jian-xun*, GUAN Gui-quan*

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Parasitology of Gansu Province/Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2017-02-10 Online:2017-07-23 Published:2017-07-23

摘要/Abstract

摘要：

本研究拟解析我国羊巴贝斯虫trap基因的结构特征，并以其为分子标志开展遗传进化分析，为理清我国羊巴贝斯虫的分类关系提供数据。用公布的巴贝斯虫trap基因序列BLAST两个羊巴贝斯虫全基因组本地数据库，以获得的保守序列片段为模板，设计PCR引物；克隆我国分离的6株羊巴贝斯虫trap基因全长序列，分析其序列结构特征；比对序列相似性，构建系统发生树，分析其遗传进化关系。结果克隆获得6株羊巴贝斯虫trap基因，分析结果显示，其具有3、4、5个内含子的三种内含子数目，预测的开放阅读框（ORF）大小分别为1 944、2 100/2 142和2 286 bp；氨基酸序列分析显示，其都具有TRAP蛋白家族特征性的vWFA、TSR、跨膜域和CTD结构域；进化关系分析显示，感染我国羊的巴贝斯虫被分成3组，组内序列相似性为99.8%~100%，而组间为43.6%~74.6%。系统发生树上6株羊巴贝斯虫被分到两大枝，羊巴贝斯虫未定种和莫氏巴贝斯虫，而且莫氏巴贝斯虫又被分成两小枝。6株羊巴贝斯虫trap基因结构存在差异，但其蛋白特征性的结构域较为保守；我国分离的羊巴贝斯虫为两个种，但莫氏巴贝斯虫中可能存在两个亚种。

Abstract:

This study focused on the structural characteristics of Chinese ovine Babesia trap gene via sequencing the full-length sequence and bioinformatics analysis. And then a phylogenetic analysis was performed on the TRAP nucleotide and amino acid sequences, which provided the data for clarifying the taxonomic relationship of Chinese ovine Babesia. The conserved sequence fragments were obtained using published Babesia trap gene sequences to BLAST 2 of local genomic databases of Chinese ovine Babesia strains. The primers of PCR were designed with the fragment sequences as templates. The full-length trap gene sequences of 6 Chinese ovine Babesia strains were cloned and their structural characteristics were analyzed. The evolutionary relationship of these ovine Babesia strains was analyzed based on the comparisons of TRAP sequence similarities and constructed phylogenetic trees. The full-length gene sequences of trap were cloned from 6 ovine Babesia strains. The results of structural characteristics analysis showed that the trap gene of 6 ovine Babesia strains had 3 patterns of intron, 3, 4 and 5 introns and the predicted ORF sizes were 1 944, 2 100/2 142 and 2 286 bp, respectively. The analysis of amino acid sequences indicated that the predicted ovine Babesia TRAPs possessed all the distinctive domains of TRAP protein family, such as vWFA, TSR, transmembrane motif and CTD. Phylogenetic analysis showed that these Chinese ovine Babesia strains could be divided into 3 groups. The similarities of intra-group between the TRAP sequences were between 99.8% and 100%, and those of inter-group were from 43.6% to 74.6%. The 6 ovine Chinese Babesia strains were divided into 2 clades, Babesia sp. and B. motasi clades. In addition, the B. motasi clade were further divided into 2 sub-clades. The trap gene structure of 6 ovine Babesia strains are various, but the distinctive protein domains are conserved. These Chinese ovine Babesia strains belong to 2 Babesia species, and possibly there are 2 sub-species in the B. motasi species.

中图分类号:

S852.723

何欣, 刘军龙, 刘爱红, 王锦明, 牛庆丽, 李有全, 殷宏, 罗建勋, 关贵全. 我国羊巴贝斯虫trap基因结构及遗传进化分析[J]. 畜牧兽医学报, 2017, 48(7): 1332-1341.

HE Xin, LIU Jun-long, LIU Ai-hong, WANG Jin-ming, NIU Qing-li, LI You-quan, YIN Hong, LUO Jian-xun, GUAN Gui-quan. The Structural and Phylogenetic Analysis of trap Gene in Ovine Babesia Species in China[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(7): 1332-1341.

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我国羊巴贝斯虫trap基因结构及遗传进化分析

The Structural and Phylogenetic Analysis of trap Gene in Ovine Babesia Species in China

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