Monastyrskaya Galina, Fushan Alexey, Abaev Igor, Filyukova Olga, Kostina Maria, Pecherskih Emily, Sverdlov Eugene
Laboratory of Structure and Functions of Human Genes, M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow V-437, Russia.
Res Microbiol. 2004 Nov;155(9):781-93. doi: 10.1016/j.resmic.2004.05.014.
Burkholderia mallei and B. pseudomallei, closely related Gram-negative bacteria, are causative agents of serious infectious diseases of humans and animals: glanders and melioidosis, respectively. Despite numerous studies of these pathogens, the detailed mechanism of their pathogenesis is still unknown. The problem is even more complicated due to natural variability of B. pseudomallei and B. mallei strains, the understanding of which is a prerequisite for rational design of tools for diagnostics, prophylaxis and therapy of the diseases. Using a subtractive hybridization technique, we compared the genomes of B. pseudomallei C-141 and B. mallei C-5 strains. A subtracted library of DNA fragments specific for B. pseudomallei C-141 and absent from B. mallei C-5 was obtained and analyzed. A variety of differences have been detected and mapped on the recently sequenced genome of B. pseudomallei K96243. A comparative sequence analysis also revealed considerable genomic differences between B. pseudomallei C-141 and B. mallei ATCC 23344 strains sequenced at The Institute for Genomic Research (TIGR). We also observed significant genomic differences between B. pseudomallei C-141 and B. pseudomallei K96243. Some of the differential DNA fragments displayed similarity to different mobile elements which have not yet been described for B. pseudomallei, whereas the others matched various prophage components, components of active transport systems, different enzymes and transcription regulators. A substantial proportion of the differential clones had no database matches either at the nucleotide or protein level. The results provide evidence for great genome-wide variability of B. pseudomallei, further confirmed by Southern blot analysis of various B. pseudomallei strains. The data obtained can be useful for future development of efficient diagnostic tools allowing rapid identification of species, strains and isolates of B. mallei and B. pseudomallei.
鼻疽伯克霍尔德菌和类鼻疽伯克霍尔德菌是密切相关的革兰氏阴性菌，分别是人类和动物严重传染病——鼻疽和类鼻疽的病原体。尽管对这些病原体进行了大量研究，但其发病机制的详细情况仍不清楚。由于类鼻疽伯克霍尔德菌和鼻疽伯克霍尔德菌菌株的自然变异性，这个问题变得更加复杂，了解这种变异性是合理设计疾病诊断、预防和治疗工具的前提。我们使用消减杂交技术比较了类鼻疽伯克霍尔德菌C-141和鼻疽伯克霍尔德菌C-5菌株的基因组。获得并分析了一个类鼻疽伯克霍尔德菌C-141特有的、鼻疽伯克霍尔德菌C-5中不存在的DNA片段消减文库。检测到了多种差异，并将其定位到最近测序的类鼻疽伯克霍尔德菌K96243基因组上。比较序列分析还揭示了类鼻疽伯克霍尔德菌C-141与美国国立基因组研究所（TIGR）测序的鼻疽伯克霍尔德菌ATCC 23344菌株之间存在相当大的基因组差异。我们还观察到类鼻疽伯克霍尔德菌C-141和类鼻疽伯克霍尔德菌K96243之间存在显著的基因组差异。一些差异DNA片段与尚未在类鼻疽伯克霍尔德菌中描述的不同移动元件相似，而其他片段则与各种噬菌体成分、主动运输系统成分、不同的酶和转录调节因子匹配。相当一部分差异克隆在核苷酸或蛋白质水平上都没有与数据库匹配的结果。这些结果为类鼻疽伯克霍尔德菌在全基因组范围内的巨大变异性提供了证据，各种类鼻疽伯克霍尔德菌菌株的Southern印迹分析进一步证实了这一点。所获得的数据可能有助于未来开发高效的诊断工具，以便快速鉴定鼻疽伯克霍尔德菌和类鼻疽伯克霍尔德菌的物种、菌株和分离株。
