Martinetti Lucchini G, Altwegg M
Institute of Medical Microbiology, University of Zürich, Switzerland.
Int J Syst Bacteriol. 1992 Jul;42(3):384-9. doi: 10.1099/00207713-42-3-384.
In the genus Aeromonas there are at least 13 DNA hybridization groups, which are difficult to differentiate biochemically. We investigated the usefulness of rRNA gene restriction patterns for characterization and identification of the various groups. Genomic DNA was digested with restriction endonuclease SmaI, transferred to a nylon membrane, and hybridized with biotinylated plasmid pKK3535 containing the rrnB operon of Escherichia coli. The SmaI bands at 0.8 to 4 kb but not those at positions corresponding to sizes larger than 4 kb showed a good correlation with hybridization groups, allowing identification of strains to the level of genetic species. We demonstrated that the 567-bp fragment localized between positions 80 and 647 of the 16S ribosomal gene of E. coli was essential for hybridization to the low-molecular-weight fragments, whereas the remainder of the operon did not hybridize to these fragments. On the basis of these results, we concluded that the Aeromonas chromosome contains multiple rRNA operons which may be used for species identification.
在气单胞菌属中,至少有13个DNA杂交组,很难通过生化方法进行区分。我们研究了rRNA基因限制性酶切图谱在各种组的特征鉴定和识别中的实用性。用限制性内切酶SmaI消化基因组DNA,转移到尼龙膜上,并用含有大肠杆菌rrnB操纵子的生物素化质粒pKK3535进行杂交。0.8至4 kb处的SmaI条带与杂交组显示出良好的相关性,而大于4 kb处的条带则不然,这使得菌株能够鉴定到遗传种水平。我们证明,位于大肠杆菌16S核糖体基因80至647位之间的567 bp片段对于与低分子量片段杂交至关重要,而操纵子的其余部分则不与这些片段杂交。基于这些结果,我们得出结论,气单胞菌染色体包含多个rRNA操纵子,可用于物种鉴定。