Multilocus sequence typing system for the endosymbiont Wolbachia pipientis
Multilocus sequence typing system for the endosymbiont Wolbachia pipientis
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DOI：
10.1128/aem.00731-06
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发表时间：
2006-11-01
影响因子：
4.4
通讯作者：
Werren, John H.
中科院分区：
文献类型：
--
作者：
Baldo, Laura;Hotopp, Julie C. Dunning;Werren, John H.
真细菌属Wolbachia包括最丰富的专性胞内细菌群之一，它的宿主范围跨越节肢动物门和线虫纲。在这里，我们开发了一种多位点序列分型(MLST)方案，作为沃尔巴克氏菌的通用基因分型工具。选择了5个普遍存在的基因(gatB、coxA、hcpA、fbpA和FtsZ)的内部片段，设计了能在节肢动物中发现的主要沃尔巴克氏菌超类群以及其他不同谱系中扩增的引物。此外，还开发了一个利用沃尔巴克氏菌表面蛋白(WSP)高变区的补充分型系统。对从单一感染宿主中分离到的A、B、D、F超群的37株菌株进行了MLST和WSP鉴定。每个MLST座位的等位基因数在25~31个之间，平均遗传多样性水平在6.5%~9.2%之间。共发现35个独特的等位基因图谱。这一结果证实了染色体基因的高度重组。结果表明，MLST可有效地检测单一寄主物种内菌株间的多样性，以及鉴定在不同节肢动物寄主中发现的密切相关菌株。不同昆虫种类的菌株具有相同或相似的等位基因图谱，并导致不同的生殖表型。具有相似WSP序列的菌株可能具有非常不同的MLST等位基因图谱，反之亦然，这表明MLST方法在菌株鉴定中的重要性。MLST系统为菌株分型、群体遗传学和分子进化研究提供了一个通用和明确的工具。用于存储和组织沃尔巴克氏菌细菌和宿主信息的中央数据库可以在http://pubmist.org/wolbachia/.上访问
The eubacterial genus Wolbachia comprises one of the most abundant groups of obligate intracellular bacteria, and it has a host range that spans the phyla Arthropoda and Nematoda. Here we developed a multilocus sequence typing (MLST) scheme as a universal genotyping tool for Wolbachia. Internal fragments of five ubiquitous genes (gatB, coxA, hcpA,fbpA, and ftsZ) were chosen, and primers that amplified across the major Wolbachia supergroups found in arthropods, as well as other divergent lineages, were designed. A supplemental typing system using the hypervariable regions of the Wolbachia surface protein (WSP) was also developed. Thirty-seven strains belonging to supergroups A, B, D, and F obtained from singly infected hosts were characterized by using MLST and WSP. The number of alleles per MLST locus ranged from 25 to 31, and the average levels of genetic diversity among alleles were 6.5% to 9.2%. A total of 35 unique allelic profiles were found. The results confirmed that there is a high level of recombination in chromosomal genes. MLST was shown to be effective for detecting diversity among strains within a single host species, as well as for identifying closely related strains found in different arthropod hosts. Identical or similar allelic profiles were obtained for strains harbored by different insect species and causing distinct reproductive phenotypes. Strains with similar WSP sequences can have very different MLST allelic profiles and vice versa, indicating the importance of the MLST approach for strain identification. The MLST system provides a universal and unambiguous tool for strain typing, population genetics, and molecular evolutionary studies. The central database for storing and organizing Wolbachia bacterial and host information can be accessed at http://pubmist.org/wolbachia/.