Department of Biotechnology and Food Microbiology, Poznań University of Life Sciences, Wojska Polskiego 48, Poznań, Poland.
Lett Appl Microbiol. 2011 Mar;52(3):281-9. doi: 10.1111/j.1472-765X.2010.02999.x. Epub 2011 Jan 17.
We have developed a PCR-based assay using custom designed panel of primers which allows rapid detection of class IIa bacteriocin-coding genes. To demonstrate the applicability of the developed assay, the method was applied on 40 metagenomic DNA preparations isolated from native microbiota of Polish artisanal cheeses produced in the Tatra Mountains.
The developed assay was designed on the basis of a large scale alignment of class IIa bacteriocin-coding genes. A panel of seven primer pairs with confirmed ability to detect class IIa bacteriocin-coding sequences was obtained. The following study has revealed a superb bacteriocinogenic potential of all forty analysed cheese samples.
The majority of obtained sequences were lactic acid bacteria (LAB) related, although some sequences showed significant similarity to bacteriocin-coding sequences present in non-LAB bacteriocin producers. The results suggest that several potentially new bacteriocin-coding sequences were found.
The developed assay can be extremely helpful in establishing whether isolates from the environment of interest have a potential of synthesizing antilisterial class IIa bacteriocins. Application of the approach may represent a useful tool contributing to ecological studies looking for valuable probiotic, bacteriocinogenic microbiota developing in foods.
我们开发了一种基于 PCR 的检测方法，使用定制设计的引物面板，可快速检测 IIa 类细菌素编码基因。为了证明所开发方法的适用性，将该方法应用于从波兰塔特拉山脉传统奶酪天然菌群中分离的 40 个宏基因组 DNA 制剂。
该检测方法是基于 IIa 类细菌素编码基因的大规模比对设计的。获得了一组 7 对引物，这些引物经证实能够检测 IIa 类细菌素编码序列。随后的研究表明，所有 40 个分析的奶酪样本都具有出色的细菌素生成潜力。
获得的大多数序列与乳酸菌（LAB）相关，尽管一些序列与非 LAB 细菌素产生菌中存在的细菌素编码序列具有显著相似性。结果表明，发现了几个潜在的新细菌素编码序列。
所开发的检测方法对于确定感兴趣环境中的分离物是否具有合成抗李斯特菌 IIa 类细菌素的潜力非常有帮助。该方法的应用可能成为一种有用的工具，有助于寻找在食品中发展的有价值的益生菌、细菌素生成菌群的生态研究。
