NR ATXP
AU Uboldi,C.; Del Vecchio,I.; Foti,M.G.; Azzalin,A.; Paulis,M.; Raimondi,E.; Vaccari,G.; Agrimi,U.; Di Guardo,G.; Comincini,S.; Ferretti,L.
TI Prion-like Doppel gene (PRND) in the goat: genomic structure, cDNA, and polymorphisms.
QU Mammalian Genome 2005 Dec; 16(12): 963-71
PT journal article
AB The genomic structure of the caprine Doppel gene (PRND) was determined using the ovine sequence as a scaffold to generate PCR fragments that were aligned with a cDNA sequence obtained from testicular mRNA. The caprine gene contains two exons, 89 and >2291 bp long, separated by a 1689-bp intron. Two mRNA isoforms of 3.2 and 4.8 kb were identified in the testis, as well as the exact transcription start site by fluorescently labeled oligonucleotide extension (FLOE). Like in sheep and cattle, the open reading frame (ORF) (537 bp) lies within exon 2 and is very much conserved in sheep (99.3%) and cattle (97%). The intronic sequence is also highly conserved (95.3%) compared with sheep, with the only exception of a 47-bp insertion. The PRND ORF was sequenced in 47 healthy and 17 TSE-affected goats of the Italian Ionica breed. Seven nucleotide positions showed variation: T28C, C65T, A151G, G286A, C385G, T451C, and T528C. Five were commonly represented polymorphisms: T28C, T451C, and T528C are silent mutations at codons L10, L151, and I176, respectively, while A151G and C385G determine a T51A and L129V amino acid change, respectively. The two remaining variants, C65T and G286A, were rare, leading to the amino acid substitutions S22F and E96K, respectively. None of the polymorphisms was significantly relatable to the TSE status, and the same result was obtained by the analysis of the combined haplotypes at the five major polymorphic sites, namely, T28C, C65T, A151G, G286A, and C385G.
MH Amino Acid Sequence; Animals; Chromosome Mapping; Cloning, Molecular; DNA, Complementary; Genomics; Goats/*genetics; Introns; Molecular Sequence Data; Open Reading Frames; *Polymorphism, Genetic; Prions/chemistry/*genetics; *Promoter Regions (Genetics); Research Support, Non-U.S. Gov't; Sequence Alignment; Sequence Homology, Nucleic Acid; Transcription Initiation Site
AD Cristina Uboldi, Igor Del Vecchio, Alberto Azzalin, Marianna Paulis, Elena Raimondi, Sergio Comincini, Luca Ferretti, Dipartimento di Genetica e Microbiologia, Universita di Pavia, via Ferrata 1, 27100, Pavia, Italy; Maria Gabriella Foti, Centro Ricerche Studi Agroalimentari FPTP-CERSA, LITA, 20090 Segrate, Italy; Gabriele Vaccari, Umberto Agrimi, Department of Food Safety and Veterinary Public Health, Istituto Superiore di Sanità, Viale Regina Elena, 299, 00161 Roma, Italy; Giovanni Di Guardo, Dipartimento di Scienze Biomediche Comparate, Università di Teramo, Piazza Aldo Moro 45, 64100 Teramo, Italy
SP englisch
PO USA