Subbotina I V, Chernykh N A, Sokolova T G, Kublanov I V, Bonch-Osmolovskaia E A, Lebedinskiĭ A V
Institute of Microbiology, Russian Academy of Sciences, pr. 60-letiya Oktyabrya 7/2, Moscow, 117312 Russia.
Mikrobiologiia. 2003 May-Jun;72(3):374-82.
Based on the analysis of 16S rRNA nucleotide sequences, oligonucleotide probes were designed for the detection and identification of representatives of the genus Thermoanaerobacter. To increase the specificity level of detection, the genus Thermoanaerobacter was divided into three groups. The probe Tab 827 (5'-GCTTCCGCDYCCCACACCTA-3') detected all known representatives of the genus Thermoanaerobacter; the probe Tab_1 844 (5'-TTAACTACGGCACGRAATGCTTC-3') was specific for the first group of the species of the genus (T. wiegelii, T. siderophilus, T. sulfurophilus, T. brockii, T. kivui, T. ethanolicus, T. acetoethylicus, and T. thermohydrosulfuricus); the probe Tab_2 424 (5'-CACTAMYGGGGTTTACAACC-3') targeted the second group (T. thermocopriae, T. mathranii, and T. italicus); and the probe Tab_3 184 (5'-TC-CTCCATCAGGATGCCCTA-3') was specific for the third group (T. tengcongensis, T. yonseiensis, T. subterraneus, and Carboxydibrachium pacificum, an organism related to the genus Thermoanaerobacter according to its 16S rRNA sequence). The oligonucleotide probes were labeled with Dig-11-dUTP. Hybridization with the probes showed the affiliation with Thermoanaerobacter of several pure cultures that were morphologically similar to representatives of this genus but possessed metabolic features unusual for it (capacity for agarose hydrolysis, anaerobic oxidation of CO, growth at low pH values) or were isolated from habitats previously unknown for Thermoanaerobacter (deep-sea hydrothermal vents).
基于对16S rRNA核苷酸序列的分析,设计了寡核苷酸探针用于嗜热厌氧菌属代表菌株的检测与鉴定。为提高检测的特异性水平,嗜热厌氧菌属被分为三组。探针Tab 827(5'-GCTTCCGCDYCCCACACCTA-3')可检测嗜热厌氧菌属的所有已知代表菌株;探针Tab_1 844(5'-TTAACTACGGCACGRAATGCTTC-3')对该属第一组物种具有特异性(维氏嗜热厌氧菌、嗜铁嗜热厌氧菌、嗜硫嗜热厌氧菌、布氏嗜热厌氧菌、基氏嗜热厌氧菌、乙醇嗜热厌氧菌、乙酰乙酸嗜热厌氧菌和嗜热硫化氢嗜热厌氧菌);探针Tab_2 424(5'-CACTAMYGGGGTTTACAACC-3')针对第二组(嗜热栖热菌、马氏嗜热菌和意大利嗜热菌);探针Tab_3 184(5'-TC-CTCCATCAGGATGCCCTA-3')对第三组具有特异性(腾冲嗜热厌氧菌、延世嗜热厌氧菌、地下嗜热厌氧菌以及太平洋羧基双臂菌,根据其16S rRNA序列,该菌与嗜热厌氧菌属相关)。寡核苷酸探针用Dig-11-dUTP进行标记。与这些探针的杂交显示,几种纯培养物与嗜热厌氧菌属相关,这些纯培养物在形态上与该属代表菌株相似,但具有该属不常见的代谢特征(琼脂糖水解能力、CO的厌氧氧化、在低pH值下生长),或者是从嗜热厌氧菌属以前未知的生境(深海热液喷口)中分离得到的。