Department of Microbiology, University of Queensland, 4072, St. Lucia, Queensland, Australia.
Microb Ecol. 1991 Dec;21(1):191-8. doi: 10.1007/BF02539153.
The 16S rDNA genes of an apparently pure culture of a psychrophilic and strict barophilic bacterium (WHB 46) were studied by PCR-mediated amplification and cloning into phage M13 mp18. Sequence analysis of five individual clones revealed the presence of two different 16S rDNA types. The homology value of 90% indicates that culture WHB 46 is actually composed of two closely related species (WHB 46-1 and 46-2). Both strains are members of the γ-subdivision of proteobacteria. Analysis of a sixth clone (WHB 46-1/2) leads to the conclusion that it represents a 16S rDNA hybrid molecule assembled during the PCR reaction. This hypothesis was confirmed by secondary structure analysis of the chimeric rDNA. The appearance of such hybrid molecules point to a potential risk in studies on the diversity of bacterial populations by analysis of rDNA pattern via PCR-mediated amplification because they suggest the existence of organisms that do not actually exist in the sample investigated.
通过 PCR 介导的扩增和克隆到噬菌体 M13mp18 中，研究了一种明显的低温和严格嗜压细菌（WHB46）的 16S rDNA 基因。对五个单独克隆的序列分析表明存在两种不同的 16S rDNA 类型。90%的同源性表明，培养物 WHB46 实际上由两个密切相关的物种（WHB46-1 和 46-2）组成。两种菌株都是γ亚群变形菌的成员。对第六个克隆（WHB46-1/2）的分析得出结论，它代表了 PCR 反应过程中组装的 16S rDNA 杂交分子。这种假设通过对嵌合 rDNA 的二级结构分析得到了证实。这种杂交分子的出现表明，通过 PCR 介导的扩增分析 rDNA 模式来研究细菌种群多样性存在潜在风险，因为它们表明存在实际上不存在于所研究样本中的生物。
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