Please use this identifier to cite or link to this item:
Type: Artigo de periódico
Title: A simplified subtractive hybridization protocol used to isolate DNA sequences specific to Xylella fastidiosa
Author: Ferreira, H
Neto, JR
Goncalves, ER
Rosato, YB
Abstract: A simplified protocol of subtractive hybridization based on the technique of L. M, Kunkel, A. P, Monaco, W, Middlesworth, H, D, Ochs & S, A. Latt (1985, Proc Natl Acad Sci USA, 82, 4778-4782) was used to obtain DNA sequences specific to Xylella fastidiosa isolated from diseased citrus plants. As a driver, DNA extracted from bacteria showing different degrees of relatedness was used: Xy, fastidiosa 788 isolated from another host (plum), Xanthomonas campestris pv, campestris and Burkholderia gladioli strains. A DNA fragment, f14, showing no hybridization to the driver DNA, was used as a probe specific to Xy. fastidiosa from citrus and oleander. This fragment was sequenced and the predicted protein showed 40% similarity to the central region of flagellin of Escherichia coli serotypes hi and H12, A pair of internal primers (f14-1 and f14-2) was designed for amplification of Xy. fastidiosa DNA, These primers detected Xy, fastidiosa strains isolated from citrus and oleander and yielded an amplification product of about 600 bp, They were also able to detect the bacteria in extracts from citrus plants with or without symptoms of disease. No amplification reaction was obtained using DNA extracted from other species and pathovars of Xanthomonas, Pseudomonas cichorii, Erwinia carotovora, Agrobacterium tumefaciens and phytopathogens of citrus (Xanthomonas axonopodis pv, citri) and coffee (Burkholderia andropogonis, P. cichorii, Pseudomonas syringae pv. garcae). The isolation of a DNA fragment specific to Xy, fastidiosa from citrus showed that the simplified protocol of subtractive hybridization used in this work is potentially applicable to other microorganisms.
Subject: Xylella fastidiosa
subtractive hybridization
PCR detection
Country: Inglaterra
Editor: Soc General Microbiology
Rights: fechado
Date Issue: 1999
Appears in Collections:Unicamp - Artigos e Outros Documentos

Files in This Item:
File Description SizeFormat 
WOS000082019000016.pdf1.27 MBAdobe PDFView/Open

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.