摘要
目的 了解腐生性钩端螺旋体(简称钩体)Potoc菌株的全基因组分子特征。方法 将腐生性钩体Potoc菌株培养后,应用高通量测序技术进行全基因组序列测定和比较基因组学分析,并将Potoc菌株灭活菌液与国内15种不同血清群致病性钩体代表菌株的兔血清进行免疫印迹分析。结果 腐生性钩体Potoc菌株全基因组序列全长为3 942 789 bp,共含有编码基因3685个,其中具有直系同源簇功能2052个,其平均GC含量为39.0%。比较基因组学分析发现Potoc菌株与国外已发表的腐生性钩体Ames菌株基因组核苷酸一致性高,同时也证实其基因组中并不存在O抗原生物合成基因簇(rfb)序列或类似基因。与问号基因种、波氏基因种和中间型L.licerasiae菌株存在共有基因1110个,而Potoc菌株基因组中所含有特有的基因共计2278个。免疫印迹分析结果发现Potoc菌株与致病性钩体中存在种属特异性的蛋白抗原成分。此外,基于全基因组水平的系统发育进化树显示,不同致病性钩体分为不同进化分支,具有特定进化历程。结论 获得了腐生性钩体Potoc菌株基因组的基本特征,探究了钩体属特异性抗原,揭示了不同基因种钩体基因组多样性,为后续致病性钩体的诊断和进化等研究奠定一定基础。
Objective To understand the whole-genomic characteristics of Leptospira biflexa Potoc strain.Methods After culturing Leptospira Potoc strain, a high-throughput sequencing technology was used to determine the whole-genome sequence and comparative genomics analysis were conducted. Inactive Potoc strain solution was analyzed by immunoblotting with rabbit sera from fifteen different serogroup pathogenic Leptospira representative strains.Results The results showed that whole-genome sequence of Leptospira Potoc strain was 3,942,789 bp in length. The genome contained 3685 coding genes, of which 2052 had the function of clusters of orthologous groups. The average GC content was 39.0%.Comparative genomics analysis showed that the Potoc strain had high nucleotide consistency with published L. biflexa Ames strain,but there was no O antigen biosynthesis gene cluster sequence or similar genes in its genome. There were 1110 genes in common with L.interrogens and borgpetersenii, intermediate species, while Potoc strain contained a total of 2278 unique genes. The immunoblotting analysis showed that there were species-specific protein antigen components in Potoc strain and pathogenic Leptospira. In addition, phylogenetic tree based on the whole genome level showed that different pathogenic Leptospiras were divided into different evolutionary branches with specific evolutionary history.Conclusion The basic characteristics of Leptospira biflexa Potoc strains have been obtained, and the specific antigens of Leptospira genus have been explored. Furthermore, the genome diversity of Leptospira with different gene species has been revealed,which lays a foundation for subsequent diagnosis and evolution of pathogenic Leptospira.
作者
张影
石刚
李彬
杜宗利
辛晓芳
徐颖华
ZHANG Ying;SHI Gang;LI Bin;DU Zong-li;XIN Xiao-fang;XU Ying-hua(NHC Key Laboratory of Biotechnology Product Verification Methods and Standardization,China Institute for Food and Drug Control,Beijing 102629,China)
出处
《中国医药生物技术》
2022年第6期497-502,共6页
Chinese Medicinal Biotechnology
基金
国家重点研发计划(2018YFC1603900)
国家自然科学基金(81471968)
中国食品药品检定研究院学科带头人基金(2013X4)。
