Wu Xi-Yang, Walker Mark J, Hornitzky Michael, Chin James
Immunology and Microbiology, Elizabeth Macarthur Agriculture Institute, NSW DPI, PMB 8, Camden, NSW 2570, Australia.
J Microbiol Methods. 2006 Jan;64(1):107-19. doi: 10.1016/j.mimet.2005.04.021. Epub 2005 Jun 24.
A group-specific primer pair was designed to amplify the 16S rRNA gene of representative reference strains from environmentally sourced, mesophilic aerobic spore-forming Bacillus taxa. The PCR generated a 1114 bp amplicon but did not do so with DNA extracted from 16 other Eubacterial species. When amplicons were digested with restriction enzymes AluI or TaqI, different profiles containing between 2 and 5 fragments ranging in size from 76 to 804 base pairs were seen with different Bacillus species. This procedure, known otherwise as amplified ribosomal DNA restriction analysis or ARDRA, produced unique and distinguishable patterns to differentiate between 15 ATCC reference strains (10 Bacillus, 3 Paenibacillus and 2 Brevibacillus member species) as well as 3 misidentified Bacillus probiotic strains in a commercial collection. Our simplified PCR-ARDRA protocol provides a facile method for the identification of most environmentally important species of Bacillus.
设计了一组特异性引物对，用于扩增来自环境来源的嗜温需氧芽孢杆菌类群代表性参考菌株的16S rRNA基因。聚合酶链反应（PCR）扩增出了一个1114 bp的扩增子，但从其他16种真细菌物种中提取的DNA则未扩增出该片段。当用限制性内切酶AluI或TaqI消化扩增子时，不同芽孢杆菌物种呈现出不同的图谱，包含2至5个片段，大小在76至804个碱基对之间。此程序，也就是扩增核糖体DNA限制性分析（ARDRA），产生了独特且可区分的模式，以区分15株美国典型培养物保藏中心（ATCC）参考菌株（10株芽孢杆菌、3株类芽孢杆菌和2株短芽孢杆菌成员物种）以及商业收藏中的3株被误认的芽孢杆菌益生菌株。我们简化的PCR-ARDRA方案为鉴定大多数对环境重要的芽孢杆菌物种提供了一种简便方法。
