Laboratory of Molecular Biology, School of Environmental Health Sciences, Azabu University, Sagamihara 229-8501, Japan.
BMC Microbiol. 2009 Dec 11;9:256. doi: 10.1186/1471-2180-9-256.
Identification and characterization of intervening sequences (IVSs) within 23S rRNA genes from Campylobacter organisms including atypical campylobacters were carried out using two PCR primer pairs, designed to generate helix 25 and 45 regions.
Only C. sputorum biovar sputorum LMG7975 and fecalis LMG8531, LMG8534 and LMG6728 of a total of 204 Campylobacter isolates (n = 56 C. jejuni; n = 11 C. coli; n = 33 C. fetus; n = 43 C. upsaliensis; n = 30 C. hyointestinalis; n = 4 C. sputorum biovar sputorum; n = 5 C. sputorum biovar fecalis; n = 5 C. sputorum biovar paraureolyticus; n = 10 C. concisus; n = 7 C. curvus) were shown to carry IVSs in helix 25 region. C. sputorum biovar fecalis LMG8531 and LMG8534, interestingly, carried two different kinds of the 23S rRNA genes with and without the IVS, respectively. Consequently, in a total of 265 isolates of 269, including 65 C. lari isolates examined previously, the absence of IVSs was identified in the helix 25 region. In the helix 45 region, all the C. hyointestinalis, C. sputorum and C. concisus isolates were shown not to carry any IVSs. However, the 30 of 56 C. jejuni isolates (54%), 5 of 11 C. coli (45%), 25 of 33 C. fetus (76%), 30 of 43 C. upsaliensis (70%) and 6 of 7 C. curvus (90%) were shown to carry IVSs. In C. jejuni and C. upsaliensis isolates, two different kinds of the 23S rRNA genes were also identified to occur with and without IVSs in the helix 45 region, respectively.
Secondary structure models were also constructed with all the IVSs identified in the present study. In the purified RNA fractions from the isolates which carried the 16S or 23S rRNA genes with the IVSs, no 16S or 23S rRNA was evident, respectively.
使用两个设计用于生成螺旋 25 和 45 区域的 PCR 引物对，对包括非典型弯曲杆菌在内的弯曲杆菌属 23S rRNA 基因中的内含子序列（IVSs）进行鉴定和特征描述。
在总共 204 个弯曲杆菌分离株（n = 56 个空肠弯曲杆菌；n = 11 个大肠弯曲杆菌；n = 33 个胎儿弯曲杆菌；n = 43 个乌普萨拉弯曲杆菌；n = 30 个海氏弯曲杆菌；n = 4 个唾液弯曲杆菌；n = 5 个粪便唾液弯曲杆菌；n = 5 个尿解质唾液弯曲杆菌；n = 10 个简约弯曲杆菌；n = 7 个弯杆菌）中，仅发现唾液弯曲杆菌和粪便唾液弯曲杆菌 LMG7975 和 LMG8531 在螺旋 25 区携带 IVSs。有趣的是，LMG8531 和 LMG8534 携带两种不同的 23S rRNA 基因，分别带有和不带有 IVS。因此，在总共 269 个分离株中，包括之前检查的 65 个 Lari 分离株，在螺旋 25 区鉴定到没有 IVS。在螺旋 45 区，所有海氏弯曲杆菌、唾液弯曲杆菌和简约弯曲杆菌分离株均未携带任何 IVS。然而，在 56 个空肠弯曲杆菌分离株中，有 30 个（54%）携带 IVS；在 11 个大肠弯曲杆菌分离株中，有 5 个（45%）携带 IVS；在 33 个胎儿弯曲杆菌分离株中，有 25 个（76%）携带 IVS；在 43 个乌普萨拉弯曲杆菌分离株中，有 30 个（70%）携带 IVS；在 7 个弯杆菌分离株中，有 6 个（90%）携带 IVS。在空肠弯曲杆菌和乌普萨拉弯曲杆菌分离株中，还发现了两种不同的 23S rRNA 基因，分别在螺旋 45 区带有和不带有 IVS。
还构建了本研究中所有鉴定到的 IVS 的二级结构模型。在携带 16S 或 23S rRNA 基因 IVS 的分离株的纯化 RNA 级分中，分别没有 16S 或 23S rRNA。
