Molecular characterization and evolution of an interspersed repetitive DNA family of oysters
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When genomic DNA from the European flat oyster Ostrea edulis L. was digested by BclI enzyme, a band of about 150 bp was observed in agarose gel. After cloning and sequencing this band and analysing their molecular characteristics and genomic organization by means of Southern blot, in situ hybridisation, and polymerase chain reaction (PCR) protocols, we concluded that this band is an interspersed highly repeated DNA element, which is related in sequence to the flanking regions of (CT)-microsatellite loci of the species O. edulis and Crassostrea gigas. Furthermore, we determined that this element forms part of a longer repetitive unit of 268 bp in length that, at least in some loci, is present in more than one copy. By Southern blot hybridisation and PCR amplifications—using primers designed for conserved regions of the 150-bp BclI clones of O. edulis—we determined that this repetitive DNA family is conserved in five other oyster species (O. stentina, C. angulata, C. gigas, C. ariakensis, and C. sikamea) while it is apparently absent in C. gasar. Finally, based on the analysis of the repetitive units in these oyster species, we discuss the slow degree of concerted evolution in this interspersed repetitive DNA family and its use for phylogenetic analysis.
KeywordsOyster Interspersed repetitive DNA Concerted evolution Ostrea Crassostrea FISH
This research was supported by grants from the Plan Andaluz de Investigación (Group Nos. BIO200 and BIO219) and a grant from the Consejería de Innovación, Ciencia y Empresa, Junta de Andalucía (Project No. C03-082). We are greatly indebted to Dr. José Ignacio Navas (CIFPA “Agua del Pino”, Huelva, Spain) for his help in sampling and technical assistance. We also thank our colleagues Pierre Boudry for providing us some oyster samples, and to our colleague David Nesbitt for revising our English text.
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