摘要
用牛巴贝斯虫棒状体颈部蛋白RON2基因序列BLAST我国羊感染的2种巴贝斯虫全基因组数据库,以获得的RON2基因保守序列片段为模板设计PCR引物;从6株羊巴贝斯虫基因组DNA和c DNA中扩增RON2基因全长序列,分析其基因结构特征;并通过序列比对和进化树构建,分析我国羊巴贝斯虫的遗传进化关系。结果显示,该基因无内含子,羊巴贝斯虫未定种新疆株/敦煌株(BspX J/DH)、莫氏巴贝斯虫天祝株/临潭株(BmTZ/LT)和莫氏巴贝斯虫宁县株/河北株(BmNX/HB)RON2蛋白基因ORF的大小分别为4 029、4 047和4 044 bp,编码1 345、1 349和1 348个氨基酸。RON2蛋白的分子质量约为151 ku,等电点为8.91,具有3个跨膜区,N端第1～29位氨基酸为其信号肽区域。RON2核苷酸和氨基酸序列比对分析结果显示,BspXJ/DH与BmTZ/LT和BmNX/HB的核苷酸和氨基酸序列相似性分别为67.0%～67.4%和69.2%～69.3%,BmTZ/LT和BmNX/HB间的相似性分别为92.8%～92.9%和91.1%～91.2%。系统发生树上羊巴贝斯虫未定种和莫氏巴贝斯虫分别位于2大支,莫氏巴贝斯虫又被分为2小支。该结果为开展我国羊巴贝斯虫的分类关系和RON2基因功能的研究提供了基础数据。
The conserved sequence fragments were obtained using rhoptry neck protein 2(RON2) gene of Babesia bovis to BLAST local genomic databases of two Babesia species infective to sheep and goats in China. The primers of PCR were designed using the fragment sequences as templates. The full-length RON2 gene sequences of 6 Chinese ovine Babesia strains were amplified from genomic DNAs and c DNAs,and these gene structural characteristics were analyzed. The genetic evolutionary relationship between these ovine Babesia strains was analyzed via sequence alignment and construction of phylogenetic trees. The result showed that there was not intron in the RON2 gene of these Babesia strains. The ORFs of Babesia sp.Xinjiang/Dunhuang(Bsp XJ/DH),B.motasi Tianzhu/Lintan(Bm TZ/LT) and B.motasi Ningxian/Hebei(BmNX/HB) RON2 gene were 4 029 bp,4 047 bp and 4 044 bp in length,and encoded 1 345,1 349 and 1 348 amino acids,respectively.The RON2 proteins of these ovine Babesia strains were about 151 ku in molecular weight and their isoelectric points were 8.91.This protein had three transmembrane domains. The initiative 29 amino acids in N terminal were the signal peptide.The alignment of RON2 nucleotide and amino acid sequences indicated that the similarities of Bsp XJ/DH and strains of B.motasi were 67.0%-67.4%and 69.2%-69.3%,and that between Bm TZ/LT and Bm NX/HB was 92.8%-92.9% and 91.1%-91.2%,respectively.The 6 ovine Babesia strains were located on 2 clades,and 4 strains of B.motasi were further divided into 2 sub-clades in phylogenetic tree.These results provided the foundational data for further exploring the genetic evolution relationship of these ovine Babesia strains and the function of Babesia RON2 gene.
作者
徐建林
王锦明
刘军龙
刘爱红
李有全
殷宏
罗建勋
关贵全
XU Jian-lin;WANG Jin-ming;LIU Jun-long;LIU Ai-hong;LI You-quan;YIN Hong;LUO Jian-xun;GUAN Gui-quan(State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veteruiary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute ,Chinese Academy of Agricultural Sciences ,Lanzhou 730046,China;Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses ,Yangzhou 225009,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2019年第5期551-559,共9页
Chinese Veterinary Science
基金
国家重点研发计划项目(2017YFD 0501200)
农业科技创新工程项目(CAAS-ASTIP-2016-LVRI)
关键词
羊巴贝斯虫
棒状体颈部蛋白2
基因结构
序列比对
遗传进化
ovine Babesia
rhoptry neck protein 2
gene structure
sequence alignment
genetic evolutionary
