| one publication added to basket [34993] | Molecular cloning of Yersinia ruckeri aroA gene: a useful taxonomic tool
In: Journal of Fish Diseases. Blackwell Science: Oxford; London; Edinburgh; Boston; Melbourne. ISSN 0140-7775; e-ISSN 1365-2761, more
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| Authors | | Top |
- Yugueros, J.
- Temprano, A.
- Luengo, J.M.
- Naharro, G.
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| Abstract |
The aroA gene of Yersinia ruckeri, which encodes 5-enolpyruvylshikimate 3-phosphate (EPSP) synthase was cloned by complementation of the aroA mutation in Escherichia coli AB2829 by using pUC18 plasmid as a vector. Nucleotide sequence of the aroA gene revealed an open reading frame of 427 amino acids showing a high degree of homology to other bacterial AroA proteins. A pair of primers with 23 and 20 nucleotides were selected from the 5’ and 3’ termini, respectively, and formed the basis of a specific polymerase chain reaction (PCR) assay. A 1165-bp deoxyribonucleic acid (DNA) fragment was amplified from all lysed Y. ruckeri strains. An identical size fragment was also amplified from lysed Y. pseudotuberculosis, Y. aldovae, Salmonella enteritidis and E. coli, but not from other enterobacteria. AluI restriction fragment length polymorphism (RFLP) of the PCR amplified products allowed for differentiation between Y. ruckeri and the other bacteria. Specificity and sensitivity make this PCR assay a useful method for rapid identification and diagnosis of Y. ruckeri infections. |
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