Systems Biology Initiative, School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Sydney, New South Wales, Australia.
PLoS One. 2011;6(7):e22170. doi: 10.1371/journal.pone.0022170. Epub 2011 Jul 29.
Campylobacter concisus is an emerging pathogen of the human gastrointestinal tract. Its role in different diseases remains a subject of debate; this may be due to strain to strain genetic variation. Here, we sequence and analyze the genome of a C. concisus from a biopsy of a child with Crohn's disease (UNSWCD); the second such genome for this species. A 1.8 Mb genome was assembled with paired-end reads from a next-generation sequencer. This genome is smaller than the 2.1 Mb C. concisus reference BAA-1457. While 1593 genes were conserved across UNSWCD and BAA-1457, 138 genes from UNSWCD and 281 from BAA-1457 were unique when compared against the other. To further validate the genome assembly and annotation, comprehensive shotgun proteomics was performed. This confirmed 78% of open reading frames in UNSWCD and, importantly, provided evidence of expression for 217 proteins previously defined as 'hypothetical' in Campylobacter. Substantial functional differences were observed between the UNSWCD and the reference strain. Enrichment analysis revealed differences in membrane proteins, response to stimulus, molecular transport and electron carriers. Synteny maps for the 281 genes not present in UNSWCD identified seven functionally associated gene clusters. These included one associated with the CRISPR family and another which encoded multiple restriction endonucleases; these genes are all involved in resistance to phage attack. Many of the observed differences are consistent with UNSWCD having adapted to greater surface interaction with host cells, as opposed to BAA-1457 which may prefer a free-living environment.
短小弯曲杆菌是人类胃肠道新兴的病原体。其在不同疾病中的作用仍然存在争议;这可能是由于菌株间遗传变异。在这里,我们对来自患有克罗恩病(UNSWCD)儿童活检的短小弯曲杆菌进行了测序和分析;这是该物种的第二个基因组。使用来自下一代测序仪的配对末端读数组装了 1.8 Mb 基因组。这个基因组比 2.1 Mb 的短小弯曲杆菌参考株 BAA-1457 小。虽然 UNSWCD 和 BAA-1457 之间有 1593 个基因保守,但与其他基因相比,UNSWCD 有 138 个基因和 BAA-1457 有 281 个基因是独特的。为了进一步验证基因组组装和注释,进行了全面的 shotgun 蛋白质组学分析。这证实了 UNSWCD 中 78%的开放阅读框,并且重要的是,为先前在弯曲杆菌中定义为“假设”的 217 种蛋白质的表达提供了证据。UNSWCD 和参考菌株之间观察到了大量的功能差异。富集分析显示,在膜蛋白、对刺激的反应、分子运输和电子载体方面存在差异。在 UNSWCD 中不存在的 281 个基因的同线性图谱确定了七个功能相关的基因簇。其中一个与 CRISPR 家族相关,另一个则编码多个限制内切酶;这些基因都参与了对噬菌体攻击的抵抗。观察到的许多差异与 UNSWCD 适应与宿主细胞更大的表面相互作用一致,而 BAA-1457 可能更喜欢自由生活的环境。