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通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和rRNA靶向寡核苷酸探针杂交对嗜酸乳杆菌、加氏乳杆菌和约翰逊乳杆菌菌株进行鉴定和分类。

Identification and classification of Lactobacillus acidophilus, L. gasseri and L. johnsonii strains by SDS-PAGE and rRNA-targeted oligonucleotide probe hybridization.

作者信息

Pot B, Hertel C, Ludwig W, Descheemaeker P, Kersters K, Schleifer K H

机构信息

Laboratorium voor Microbiologie, Universiteit Gent, Belgium.

出版信息

J Gen Microbiol. 1993 Mar;139(3):513-7. doi: 10.1099/00221287-139-3-513.

DOI:10.1099/00221287-139-3-513
PMID:7682599
Abstract

Thirty-two strains originally identified as Lactobacillus acidophilus and L. gasseri were screened for their taxonomic homogeneity by SDS-PAGE of whole-cell proteins. After numerical comparison of the resulting protein electrophoretic fingerprints, two well-delineated clusters were detected. The majority of the strains grouped in one electrophoretic cluster, which contained the type strain of L. acidophilus and corresponds to DNA group A1 of Johnson, J. L., Phelps, C. F., Cummins, C. S., London, J. & Gasser, F. (1980; International Journal of Systematic Bacteriology 30, 53-68). Another cluster corresponded to DNA group B. It contained two subclusters, which agreed perfectly with DNA subgroups B1 (L. gasseri) and B2 (L. johnsonii), respectively. The 23S rRNA genes were partially sequenced and 23S-rRNA-targeted oligonucleotide probes were designed for identification of DNA groups A1, B1 and B2. Probe Lbg reacted with all strains of electrophoretic cluster B1 (L. gasseri), probe Lbj hybridized with strains of cluster B2 (L. johnsonii) and probe Lba with strains of cluster A1 (authentic L. acidophilus). The probes were successfully used for the identification of strains belonging to the respective species. The phylogenetic relationship of a representative of L. johnsonii was determined by comparative sequence analysis of the 16S rRNA genes. It is very closely related to L. gasseri.

摘要

通过全细胞蛋白的SDS-PAGE对最初鉴定为嗜酸乳杆菌和格氏乳杆菌的32株菌株进行分类学同质性筛选。对所得蛋白质电泳指纹图谱进行数值比较后,检测到两个界限分明的聚类。大多数菌株聚集在一个电泳聚类中,该聚类包含嗜酸乳杆菌的模式菌株,对应于Johnson, J. L., Phelps, C. F., Cummins, C. S., London, J. & Gasser, F.(1980年;《国际系统细菌学杂志》30, 53 - 68)的DNA组A1。另一个聚类对应于DNA组B。它包含两个亚聚类,分别与DNA亚组B1(格氏乳杆菌)和B2(约氏乳杆菌)完全一致。对23S rRNA基因进行了部分测序,并设计了针对23S rRNA的寡核苷酸探针用于鉴定DNA组A1、B1和B2。探针Lbg与电泳聚类B1(格氏乳杆菌)的所有菌株反应,探针Lbj与聚类B2(约氏乳杆菌)的菌株杂交,探针Lba与聚类A1(正宗嗜酸乳杆菌)的菌株杂交。这些探针已成功用于鉴定属于各自物种的菌株。通过对16S rRNA基因的比较序列分析确定了约氏乳杆菌一个代表菌株的系统发育关系。它与格氏乳杆菌密切相关。

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