Poly Frédéric, Threadgill Deborah, Stintzi Alain
Department of Veterinary Pathobiology, College of Veterinary Medicine, Oklahoma State University, Stillwater, OK 74078, USA.
J Bacteriol. 2004 Jul;186(14):4781-95. doi: 10.1128/JB.186.14.4781-4795.2004.
This study describes a novel approach to identify unique genomic DNA sequences from the unsequenced strain C. jejuni ATCC 43431 by comparison with the sequenced strain C. jejuni NCTC 11168. A shotgun DNA microarray was constructed by arraying 9,600 individual DNA fragments from a C. jejuni ATCC 43431 genomic library onto a glass slide. DNA fragments unique to C. jejuni ATCC 43431 were identified by competitive hybridization to the array with genomic DNA of C. jejuni NCTC 11168. The plasmids containing unique DNA fragments were sequenced, allowing the identification of up to 130 complete and incomplete genes. Potential biological roles were assigned to 66% of the unique open reading frames. The mean G+C content of these unique genes (26%) differs significantly from the G+C content of the entire C. jejuni genome (30.6%). This suggests that they may have been acquired through horizontal gene transfer from an organism with a G+C content lower than that of C. jejuni. Because the two C. jejuni strains differ by Penner serotype, a large proportion of the unique ATCC 43431 genes encode proteins involved in lipooligosaccharide and capsular biosynthesis, as expected. Several unique open reading frames encode enzymes which may contribute to genetic variability, i.e., restriction-modification systems and integrases. Interestingly, many of the unique C. jejuni ATCC 43431 genes show identity with a possible pathogenicity island from Helicobacter hepaticus and components of a potential type IV secretion system. In conclusion, this study provides a valuable resource to further investigate Campylobacter diversity and pathogenesis.
本研究描述了一种新方法,通过与已测序的空肠弯曲菌NCTC 11168菌株进行比较,从未经测序的空肠弯曲菌ATCC 43431菌株中鉴定独特的基因组DNA序列。通过将来自空肠弯曲菌ATCC 43431基因组文库的9600个单个DNA片段排列在载玻片上,构建了鸟枪法DNA微阵列。通过与空肠弯曲菌NCTC 11168的基因组DNA与该阵列进行竞争性杂交,鉴定出空肠弯曲菌ATCC 43431特有的DNA片段。对含有独特DNA片段的质粒进行测序,从而鉴定出多达130个完整和不完整的基因。66%的独特开放阅读框被赋予了潜在的生物学功能。这些独特基因的平均G+C含量(26%)与整个空肠弯曲菌基因组的G+C含量(30.6%)有显著差异。这表明它们可能是通过水平基因转移从G+C含量低于空肠弯曲菌的生物体中获得的。由于这两种空肠弯曲菌菌株在Penner血清型上存在差异,正如预期的那样,空肠弯曲菌ATCC 43431的大部分独特基因编码参与脂寡糖和荚膜生物合成的蛋白质。几个独特的开放阅读框编码可能导致遗传变异性的酶,即限制修饰系统和整合酶。有趣的是,空肠弯曲菌ATCC 43431的许多独特基因与来自肝螺杆菌的一个可能的致病岛和一个潜在的IV型分泌系统的成分具有同源性。总之,本研究为进一步研究弯曲杆菌的多样性和发病机制提供了宝贵的资源。