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Genomic DNA cloning of rickettsia-like organisms (RLO) of Saint-Jacques scallop Pecten maximus : evaluation of prokaryote diagnosis by hybridization with a non-isotopically labelled probe and by polymerase chain reaction ArchiMer
Kellner-cousin, Kristell; Le Gall, G; Despres, Béatrice; Kaghad, M; Legoux, P; Shire, D; Mialhe, Eric.
Genomic DNA fragments of a rickettsia-like organism (RLO) of Saint-Jacques scallop, Pecten maximus, were extracted from purified RLO and cloned. A DNA sequence of 1500 bp taken from one of the sequenced clones was coupled to horseradish peroxidase and the resulting probe was assayed for the diagnosis of RLO by nucleic acid hybridization. The specificity of the probe for RLO DNA was established from its lack of hybridization with scallop DNA. The sensitivity limit of the probe detected by enhanced chemiluminescence was determined to be around 500 ng of total RLO nucleic acids, equivalent to about 2.5 X 106 copies of RLO DNA. A pair of polymerase chain reaction (PCR) primers were synthesized that amplified a 1300 bp segment of the 1500 bp RLO fragment. After...
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Ano: 1993 URL: http://archimer.ifremer.fr/doc/1993/publication-2771.pdf
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