Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5907634 | Gene | 2008 | 9 Pages |
Abstract
We have sequenced and partially characterized an Echinococcus granulosus cDNA, termed egat1, from a protoscolex signal sequence trap (SST) cDNA library. The isolated 1627Â bp long cDNA contains an ORF of 489 amino acids and shows an amino acid identity of 30% with neutral and excitatory amino acid transporters members of the Dicarboxylate/Amino Acid Na+ and/or H+ Cation Symporter family (DAACS) (TC 2.A.23). Additional bioinformatics analysis of EgAT1, confirmed the results obtained by similarity searches and showed the presence of 9 to 10 transmembrane domains, consensus sequences for N-glycosylation between the third and fourth transmembrane domain, a highly similar hydropathy profile with ASCT1 (a known member of DAACS family), high score with SDF (Sodium Dicarboxilate Family) and similar motifs with EDTRANSPORT, a fingerprint of excitatory amino acid transporters. The localization of the putative amino acid transporter was analyzed by in situ hybridization and immunofluorescence in protoscoleces and associated germinal layer. The in situ hybridization labelling indicates the distribution of egat1 mRNA throughout the tegument. EgAT1 protein, which showed in Western blots a molecular mass of ~Â 60Â kD, is localized in the subtegumental region of the metacestode, particularly around suckers and rostellum of protoscoleces and layers from brood capsules. The sequence and expression analyses of EgAT1 pave the way for functional analysis of amino acids transporters of E. granulosus and its evaluation as new drug targets against cystic echinococcosis.
Keywords
ASCTBCIPNBTSSTPBSTSDFSSCORFTween-20TrxPBSBSAcDNADNA complementary to RNAbovine serum albuminHelminth parasitesCystic echinococcosisnitro blue tetrazoliumReverse transcriptaseELISAEnzyme-linked immunosorbent assayrapid amplification of cDNA endsSignal sequence trapsodium saline citrateopen reading framePhosphate-buffered salineRaceUTR یا untranslated regions untranslated regionpolymerase chain reactionPCR
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Authors
Federico Camicia, Rodolfo Paredes, Cora Chalar, Norbel Galanti, Laura Kamenetzky, Ariana Gutierrez, Mara C. Rosenzvit,