Parker Craig T, Quiñones Beatriz, Miller William G, Horn Sharon T, Mandrell Robert E
U.S. Department of Agriculture, Agricultural Research Service, Western Regional Research Center, Produce Safety and Microbiology Unit, 800 Buchanan Street, Albany, CA 94710, USA.
J Clin Microbiol. 2006 Nov;44(11):4125-35. doi: 10.1128/JCM.01231-06. Epub 2006 Aug 30.
Analysis of the complete genomic sequence of Campylobacter jejuni strain RM1221 identified four large genomic elements, Campylobacter jejuni-integrated elements (CJIEs), that were absent from C. jejuni strain NCTC 11168. To further investigate the genomic diversity of Campylobacter, we conducted a comparative genomic analysis from a collection of 67 C. jejuni and 12 Campylobacter coli strains isolated from various geographical locations and clinical and veterinary sources. Utilizing PCR, we demonstrated that 55% of the C. jejuni strains examined were positive for at least one RM1221-like genomic element and 27% were positive for two or more of these CJIEs. Furthermore, many C. coli strains were positive for either genomic element CJIE1 or CJIE3. To simultaneously assess for the presence or absence of several genes that comprise the various CJIEs, we developed a multistrain C. jejuni DNA microarray that contained most of the putative coding sequences for strains NCTC 11168 and RM1221. A comparative genomic hybridization (CGH) analysis of 35 of the 67 C. jejuni strains confirmed the presence of genomic elements similar to those in strain RM1221. Interestingly, the DNA microarray analysis demonstrated that these genomic elements in the other C. jejuni strains often exhibited modular patterns with some regions of the CJIEs present and other regions either absent or highly divergent compared to strain RM1221. Our CGH method also identified 18 other intraspecies hypervariable regions, such as the capsule and lipooligosaccharide biosynthesis regions. Thus, the inclusion of genes from these integrated genomic elements and the genes from the other intraspecies hypervariable regions contributes to a better assessment of the diversity in C. jejuni and may increase the usefulness of DNA microarrays as an epidemiological genotyping tool. Finally, we also showed that in CJIE1, a Campylobacter Mu-like phage, is located differentially in other strains of C. jejuni, suggesting that it may integrate essentially randomly.
空肠弯曲菌RM1221菌株的全基因组序列分析鉴定出四个大型基因组元件,即空肠弯曲菌整合元件(CJIEs),这些元件在空肠弯曲菌NCTC 11168菌株中不存在。为了进一步研究弯曲菌的基因组多样性,我们对从不同地理位置以及临床和兽医来源分离的67株空肠弯曲菌和12株结肠弯曲菌菌株进行了比较基因组分析。利用聚合酶链反应(PCR),我们证明,所检测的55%的空肠弯曲菌菌株至少对一种类似RM1221的基因组元件呈阳性,27%的菌株对两种或更多种这些CJIEs呈阳性。此外,许多结肠弯曲菌菌株对基因组元件CJIE1或CJIE3呈阳性。为了同时评估构成各种CJIEs的几个基因的有无,我们开发了一种多菌株空肠弯曲菌DNA微阵列,其中包含NCTC 11168和RMl221菌株的大部分推定编码序列。对67株空肠弯曲菌菌株中的35株进行的比较基因组杂交(CGH)分析证实存在与RM1221菌株中类似的基因组元件。有趣的是,DNA微阵列分析表明,其他空肠弯曲菌菌株中的这些基因组元件通常呈现模块化模式,与RM1221菌株相比,CJIEs的某些区域存在,而其他区域要么不存在,要么高度 divergent 。我们的CGH方法还鉴定出18个其他种内高变区,如荚膜和脂寡糖生物合成区。因此,纳入这些整合基因组元件的基因以及其他种内高变区的基因有助于更好地评估空肠弯曲菌的多样性,并可能增加DNA微阵列作为一种流行病学基因分型工具的实用性。最后,我们还表明,在CJIE1中,一种类似弯曲菌Mu的噬菌体在其他空肠弯曲菌菌株中的定位不同,这表明它可能基本上是随机整合的。