Kaleta Pawel, Callanan Michael J, O'Callaghan John, Fitzgerald Gerald F, Beresford Thomas P, Ross R Paul
Teagasc, Moorepark Food Research Centre, Moorepark, Fermoy, Co. Cork, Ireland.
J Microbiol Methods. 2009 Oct;79(1):32-6. doi: 10.1016/j.mimet.2009.07.019. Epub 2009 Jul 29.
The species Lactobacillus helveticus is a commonly used thermophilic starter and/or adjunct culture for Swiss and Cheddar cheese manufacture. Its use is normally associated with flavour improvement which is known to be associated with culture traits such as rapid autolysis and high proteolytic activity. The genome of the commercial strain, DPC4571, was recently sequenced and found to have an abundance of IS sequences in terms of both abundance (213 intact) and diversity (21 types). Given this unique diversity for a lactic acid bacterium, we investigated whether PCR-based IS fingerprinting could be used as a discriminatory tool to distinguish between different strains of Lb. helveticus. A set of ten primers targeting five of the most numerous groups (ISL1201, ISLhe65, ISLhe2, ISLhe15 and ISL2) of IS elements was designed. Multiplex-PCR with all primers resulted in 1-12 discreet amplicons for each strain tested. The resultant fingerprints (in the 0.5 kb-3 kb range) were found to be strain specific and reproducible. This approach thus provides a valuable method to distinguish between Lb. helveticus strains while giving some indication of the relative abundance of IS sequences in each strain.
瑞士乳杆菌是用于生产瑞士奶酪和切达干酪的常用嗜热发酵剂和/或辅助培养物。其使用通常与风味改善有关,已知风味改善与诸如快速自溶和高蛋白水解活性等培养特性相关。商业菌株DPC4571的基因组最近已被测序,发现其在丰度(213个完整)和多样性(21种类型)方面都有大量的IS序列。鉴于乳酸菌具有这种独特的多样性,我们研究了基于PCR的IS指纹图谱是否可以用作区分不同瑞士乳杆菌菌株的鉴别工具。设计了一组针对五个数量最多的IS元件组(ISL1201、ISLhe65、ISLhe2、ISLhe15和ISL2)的十种引物。使用所有引物进行多重PCR,每个测试菌株产生1至12个离散的扩增子。发现所得指纹图谱(在0.5 kb至3 kb范围内)具有菌株特异性且可重复。因此,这种方法提供了一种区分瑞士乳杆菌菌株的有价值的方法,同时还能给出每个菌株中IS序列相对丰度的一些指示。