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将气单胞菌属aroA基因的限制性片段长度多态性聚合酶链反应(RFLP-PCR)分析作为一种分类工具。

RFLP-PCR analysis of the aroA gene as a taxonomic tool for the genus Aeromonas.

作者信息

Cascón Soriano A, Anguita Castillo J, Hernanz Moral C, Sánchez Salazar M, Yugueros Marcos J, Naharro Carrasco G

机构信息

Departamento de Patología Animal, Sanidad Animal, Microbiología e Inmunología, Facultad de Veterinaria, Universidad de León, Spain.

出版信息

FEMS Microbiol Lett. 1997 Nov 15;156(2):199-204. doi: 10.1111/j.1574-6968.1997.tb12727.x.

Abstract

The aroA gene has been identified as a target in screening for the presence of most Aeromonas species so far described by PCR. Synthetic oligonucleotide primers of 24 and 25 nucleotides were used by PCR assay to amplify a sequence of the aroA gene, which encodes 3-phosphoshikimate-1-carboxyvinyltransferase, a key enzyme of aromatic amino acids and folate biosynthetic pathway. A 1236-bp DNA fragment, representing most of the aroA gene, according to the nucleotide sequence of A. salmonicida, was amplified from all Aeromonas species tested, which represented most of the 14 hybridization groups. HaeII digestion of the 1236-bp fragment generated a restriction fragment length polymorphisms which could be used as a powerful tool for identification of aeromonads to the genus level.

摘要

aroA基因已被确定为通过PCR筛选迄今所描述的大多数气单胞菌属物种时的一个靶标。PCR检测使用了24和25个核苷酸的合成寡核苷酸引物来扩增aroA基因的一个序列,该基因编码3-磷酸莽草酸-1-羧乙烯基转移酶,这是芳香族氨基酸和叶酸生物合成途径的一种关键酶。根据杀鲑气单胞菌的核苷酸序列,从所有测试的气单胞菌属物种中扩增出一个代表aroA基因大部分的1236 bp DNA片段,这些物种代表了14个杂交组中的大多数。对1236 bp片段进行HaeII消化产生了限制性片段长度多态性,可作为在属水平鉴定气单胞菌的有力工具。

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