Usui Kaori, Iida Haruna, Ueno Hitomi, Sekizuka Tsuyoshi, Matsuda Motoo, Murayama Ohoshi, Cherie Millar B, Moore John E
Laboratory of Molecular Biology, School of Environmental Health Sciences, Azabu University, Fuchinobe 1-17-71, Sagamihara 229-8501, Japan.
Int J Hyg Environ Health. 2006 Nov;209(6):541-5. doi: 10.1016/j.ijheh.2005.09.010. Epub 2006 Jun 23.
Degenerate PCR primers in silico based on the two urease structural genes, ureA and ureB, were designed for urease-positive thermophilic Campylobacter (UPTC). Resultant PCR amplification employing these primers generated an amplicon of approximately 2kb, which was cloned and sequenced in UPTC (n=12) isolated from various parts of Europe and Japan. Overall, sequence similarities were shown to be 96.7 to 99.9%. Following sequence alignment analysis, the approximate 1.96kb regions were deduced to consist of parts of ureA (about 570bps) and ureB (about 1390bps) with an overlapping region between the ureA and ureB gene loci. Although a total of 144 heterogeneous sites of all substitutions were located throughout this region, the substitution ratio was higher in the ureA region (1/Omega10bases) than in the ureB region (1/Omega15bases). A resulting dendrogram was constructed, which was based on the nucleotide sequence data of 12 UPTC isolates and demonstrated that the UPTC were genetically variable. They formed a major cluster with Helicobacter, separate from the other urease-producing bacteria examined, suggesting a shared ancestry between UPTC and Helicobacter.
基于脲酶结构基因ureA和ureB，通过计算机设计了简并PCR引物，用于脲酶阳性嗜热弯曲菌（UPTC）。使用这些引物进行PCR扩增，产生了一个约2kb的扩增子，该扩增子在从欧洲和日本各地分离的UPTC（n = 12）中进行了克隆和测序。总体而言，序列相似性显示为96.7%至99.9%。经过序列比对分析，推断约1.96kb的区域由ureA的部分（约570bp）和ureB的部分（约1390bp）组成，ureA和ureB基因座之间存在重叠区域。尽管在该区域共发现了144个所有替换的异质位点，但ureA区域的替换率（1/每10个碱基）高于ureB区域（1/每15个碱基）。构建了一个基于12个UPTC分离株核苷酸序列数据的系统发育树，结果表明UPTC具有遗传变异性。它们与幽门螺杆菌形成一个主要聚类，与其他检测的产脲酶细菌分开，这表明UPTC和幽门螺杆菌之间有共同的祖先。
