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基于16S-23S rRNA基因间隔区的异质性对突尼斯发酵肉制品中乳酸菌进行的多相分类研究。

Polyphasic taxonomic studies of lactic acid bacteria associated with Tunisian fermented meat based on the heterogeneity of the 16S-23S rRNA gene intergenic spacer region.

作者信息

Ben Belgacem Zouhaier, Dousset Xavier, Prévost Hervé, Manai Mohamed

机构信息

Faculté des Sciences de Tunis, Laboratoire de Biochimie et Biologie Moléculaire, Campus Universitaire El-Manar, 2092 Tunis, Tunisie.

出版信息

Arch Microbiol. 2009 Sep;191(9):711-20. doi: 10.1007/s00203-009-0499-2. Epub 2009 Aug 11.

Abstract

The objective of this work was to investigate the structure and diversity of lactic acid bacteria (LAB) communities in traditionally fermented meat collected from different areas of Tunisia. A polyphasic study, which involves phenotypic tests and ribosomal DNA-based techniques, was used to identify Gram-positive and catalase-negative isolates. PCR amplification of the 16S-23S rDNA ISR of 102 isolates and other reference LAB strains gave (1) one type of rrn operon (M-ISR) for lactococci, (2) two types of rrn operon (S-ISR and M-ISR) for enterococci, (3) two types of rrn operon (S-ISR and L-ISR) for Lactobacilli, and (4) three PCR amplicons (S-ISR, M-ISR, and L-ISR) obtained for Pediococcus spp. and Weissella genus. The clustering and comparison of ISR-RFLP profiles given by the isolates with those given by reference LAB strains, allowed their identification as Lactococcus lactis, Enterococcus faecium, Enterococcus faecalis, Enterococcus sanguinicola, Enterococcus hawaiiensis, Lactobacillus sakei, Lactobacillus curvatus, Lactobacillus plantarum, Lactobacillus alimentarius, Pediococcus pentosaceus, and Weissella confusa. Combined 16S-23S rDNA ISR and RFLP patterns can be considered as a good potential target for a rapid and reliable differentiation between isolates of LAB and provided further information on the organization of their rrn operons.

摘要

本研究的目的是调查从突尼斯不同地区采集的传统发酵肉类中乳酸菌(LAB)群落的结构和多样性。采用了一项多相研究,包括表型测试和基于核糖体DNA的技术,以鉴定革兰氏阳性和过氧化氢酶阴性分离株。对102株分离株和其他参考LAB菌株的16S-23S rDNA间隔序列区域(ISR)进行PCR扩增,结果显示:(1)乳酸球菌有1种rrn操纵子类型(M-ISR);(2)肠球菌有2种rrn操纵子类型(S-ISR和M-ISR);(3)乳酸杆菌有2种rrn操纵子类型(S-ISR和L-ISR);(4)嗜盐四联球菌属和魏斯氏菌属获得3种PCR扩增产物(S-ISR、M-ISR和L-ISR)。将分离株的ISR-RFLP图谱与参考LAB菌株的图谱进行聚类和比较,可将它们鉴定为乳酸乳球菌、屎肠球菌、粪肠球菌、血肠球菌、夏威夷肠球菌、清酒乳杆菌、弯曲乳杆菌、植物乳杆菌、食淀粉乳杆菌、戊糖片球菌和困惑魏斯氏菌。16S-23S rDNA ISR和RFLP联合模式可被视为快速可靠地区分LAB分离株的良好潜在靶点,并提供了有关其rrn操纵子组织的进一步信息。

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