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用于植物乳杆菌菌株快速分型的随机扩增多态性DNA(RAPD)

Randomly amplified polymorphic DNA (RAPD) for rapid typing of Lactobacillus plantarum strains.

作者信息

Johansson M L, Quednau M, Molin G, Ahrné S

机构信息

Department of Food Technology, Lund University, Sweden.

出版信息

Lett Appl Microbiol. 1995 Sep;21(3):155-9. doi: 10.1111/j.1472-765x.1995.tb01030.x.

Abstract

Randomly amplified polymorphic DNA (RAPD) has been used for rapid typing of Lactobacillus plantarum strains. RAPD was used with either purified chromosomal DNA serving as template in the polymerase chain reaction, or with crude cell extracts, and using a 9-mer primer with 80% G+C content. Amplified DNA was visualized by ethidium bromide staining after separation on agarose gels. Patterns from 20 Lact. plantarum strains and two Lact. pentosus strains were analysed using the Pearson products moment correlation coefficient (r) and the unweighted pair group method with arithmetic averages (UPGMA). With some exceptions, the two sources of template DNA gave the same clusters and subclusters of strains at the similarity level of 50%. About 50% of the strains could be individually separated from all the other tested strains. The buffer brand, the amount of primer and crude cell extract used in the PCR-step were crucial for the final pattern.

摘要

随机扩增多态性DNA(RAPD)已用于植物乳杆菌菌株的快速分型。RAPD在聚合酶链反应中使用纯化的染色体DNA作为模板,或使用粗细胞提取物,并使用G+C含量为80%的9聚体引物。扩增的DNA在琼脂糖凝胶上分离后用溴化乙锭染色进行可视化。使用皮尔逊积矩相关系数(r)和算术平均的非加权配对组方法(UPGMA)分析了20株植物乳杆菌菌株和2株戊糖乳杆菌菌株的图谱。除了一些例外情况,在50%的相似性水平上,两种模板DNA来源给出了相同的菌株簇和亚簇。约50%的菌株可以与所有其他测试菌株单独区分开来。PCR步骤中使用的缓冲液品牌、引物量和粗细胞提取物对最终图谱至关重要。

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