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. | - CCMAR -

Journal Article

TítuloDevelopment of an in vitro clonal culture and characterization of the rRNA gene cluster of Perkinsus atlanticus, a protistan parasite of the clam Tapes decussatus.
Publication TypeJournal Article
AuthorsRobledo, JAF, Nunes, PA, M. Cancela, L, Vasta, GR
Year of Publication2002
JournalJ Eukaryot Microbiol
Volume49
Questão5
Date Published2002 Sep-Oct
Pagination414-22
ISSN1066-5234
Palavras-chaveAnimals, Apicomplexa, Aquaculture, Base Sequence, Bivalvia, DNA, Protozoan, DNA, Ribosomal Spacer, Genes, rRNA, Molecular Sequence Data, Multigene Family, Polymerase Chain Reaction, RNA, Ribosomal, Sequence Analysis, DNA, Species Specificity
Abstract

Perkinsus 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.

Sapientia

http://www.ncbi.nlm.nih.gov/pubmed/12425530?dopt=Abstract

Alternate JournalJ. Eukaryot. Microbiol.
PubMed ID12425530
CCMAR Authors