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Development of an in vitro clonal culture and characterization of the rRNA gene cluster of Perkinsus atlanticus, a protistan parasite of the clam Tapes decussatus

dc.contributor.authorRobledo, J. A. F.
dc.contributor.authorNunes, Patrícia A.
dc.contributor.authorCancela, Leonor
dc.contributor.authorVasta, G. R.
dc.date.accessioned2015-11-30T15:17:51Z
dc.date.available2015-11-30T15:17:51Z
dc.date.issued2002
dc.description.abstractPerkinsus atlanticus cultures were established either with trophozoites isolated from fresh gills, with hypnospores isolated from tissues incubated in fluid thioglycollate medium, or directly from infected hemocytes of carpet shell clams Tapes decussatus from Algarve (Southern Portugal), using a culture medium and conditions optimized for Perkinsus marinus. Perkinsus atlanticus isolates were cloned by limiting dilution, and their identity unequivocally established by PCR-based species-specific diagnostic assays, and by sequencing the complete rRNA gene cluster. The rRNA gene cluster is 7.5-kb in length including 5S, IGS, SSU, ITS1, 5.8S, ITS2, LSU, and an inter-cluster spacer. rDNA sequences of the P. atlanticus clone were between 98.3-100% identical to P. atlanticus sequences previously obtained from clam tissue (non-clonal) isolates. Based on the IGS sequences available from Perkinsus species, a set of primers was designed to amplify P. atlanticus and the two clonally-cultured Perkinsus species (P. marinus and P. andrewsi) currently available from a recognized repository. This Perkinsus "genus-specific" PCR-based assay complements the species-specific assays developed earlier and strengthen the detection of Perkinsus species for which specific detection assays are not yet available.
dc.identifier.doihttps://dx.doi.org/10.1111/j.1550-7408.2002.tb00221.x
dc.identifier.issn1066-5234
dc.identifier.otherAUT: PCA01721;
dc.identifier.urihttp://hdl.handle.net/10400.1/7221
dc.language.isoeng
dc.peerreviewedyes
dc.publisherWiley
dc.relation.isbasedonP-000-N5J
dc.titleDevelopment of an in vitro clonal culture and characterization of the rRNA gene cluster of Perkinsus atlanticus, a protistan parasite of the clam Tapes decussatus
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage422
oaire.citation.startPage414
oaire.citation.titleJournal of Eukaryotic Microbiology
oaire.citation.volume49
person.familyNameNunes Cabrita
person.familyNameCancela
person.givenNamePatrícia
person.givenNameM. Leonor
person.identifier.orcid0000-0002-7148-8950
person.identifier.orcid0000-0003-3114-6662
person.identifier.scopus-author-id7006823084
rcaap.rightsrestrictedAccess
rcaap.typearticle
relation.isAuthorOfPublication4b0b5899-4a35-4405-acbe-a8f548457da1
relation.isAuthorOfPublicationb9bbfe32-3dfe-4131-ad14-a4394008447f
relation.isAuthorOfPublication.latestForDiscoveryb9bbfe32-3dfe-4131-ad14-a4394008447f

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