Department of Comparative Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, Indiana, United States of America.
PLoS One. 2011 May 10;6(5):e19574. doi: 10.1371/journal.pone.0019574.
Mycoplasma suis, the causative agent of porcine infectious anemia, has never been cultured in vitro and mechanisms by which it causes disease are poorly understood. Thus, the objective herein was to use whole genome sequencing and analysis of M. suis to define pathogenicity mechanisms and biochemical pathways. M. suis was harvested from the blood of an experimentally infected pig. Following DNA extraction and construction of a paired end library, whole-genome sequencing was performed using GS-FLX (454) and Titanium chemistry. Reads on paired-end constructs were assembled using GS De Novo Assembler and gaps closed by primer walking; assembly was validated by PFGE. Glimmer and Manatee Annotation Engine were used to predict and annotate protein-coding sequences (CDS). The M. suis genome consists of a single, 742,431 bp chromosome with low G+C content of 31.1%. A total of 844 CDS, 3 single copies, unlinked rRNA genes and 32 tRNAs were identified. Gene homologies and GC skew graph show that M. suis has a typical Mollicutes oriC. The predicted metabolic pathway is concise, showing evidence of adaptation to blood environment. M. suis is a glycolytic species, obtaining energy through sugars fermentation and ATP-synthase. The pentose-phosphate pathway, metabolism of cofactors and vitamins, pyruvate dehydrogenase and NAD(+) kinase are missing. Thus, ribose, NADH, NADPH and coenzyme A are possibly essential for its growth. M. suis can generate purines from hypoxanthine, which is secreted by RBCs, and cytidine nucleotides from uracil. Toxins orthologs were not identified. We suggest that M. suis may cause disease by scavenging and competing for host' nutrients, leading to decreased life-span of RBCs. In summary, genome analysis shows that M. suis is dependent on host cell metabolism and this characteristic is likely to be linked to its pathogenicity. The prediction of essential nutrients will aid the development of in vitro cultivation systems.
猪传染性贫血的病原体猪肺炎支原体从未在体外培养过,其致病机制也知之甚少。因此,本研究旨在利用猪肺炎支原体的全基因组测序和分析来定义其致病机制和生化途径。从实验感染猪的血液中收获猪肺炎支原体。在提取 DNA 并构建配对末端文库后,使用 GS-FLX(454)和 Titanium 化学进行全基因组测序。使用 GS De Novo 组装器组装配对末端构建物,并通过引物行走封闭缺口;通过 PFGE 验证组装。使用 Glimmer 和 Manatee Annotation Engine 预测和注释蛋白编码序列(CDS)。猪肺炎支原体基因组由一个单一的、742,431 bp 的染色体组成,GC 含量低,为 31.1%。共鉴定出 844 个 CDS、3 个单拷贝、不连锁的 rRNA 基因和 32 个 tRNA。基因同源性和 GC 倾斜图表明,猪肺炎支原体具有典型的 Mollicutes oriC。预测的代谢途径简明扼要,显示出对血液环境适应的证据。猪肺炎支原体是一种糖酵解菌,通过糖发酵和 ATP 合酶获取能量。戊糖磷酸途径、辅酶和维生素代谢、丙酮酸脱氢酶和 NAD(+)激酶缺失。因此,核糖、NADH、NADPH 和辅酶 A 可能对其生长至关重要。猪肺炎支原体可以从 RBC 分泌的次黄嘌呤和尿嘧啶中生成嘌呤和胞苷核苷酸。未鉴定到毒素同源物。我们认为,猪肺炎支原体可能通过掠夺和竞争宿主的营养物质来引起疾病,导致 RBC 寿命缩短。总之,基因组分析表明,猪肺炎支原体依赖于宿主细胞代谢,这种特性可能与其致病性有关。必需营养素的预测将有助于体外培养系统的开发。