title: Prion-like Doppel gene (PRND) in the goat: genomic structure, cDNA, and polymorphisms.
abstract: 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.
authors.affiliation: Dipartimento di Genetica e Microbiologia, Università di Pavia, via Ferrata 1, 27100, Pavia, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Uboldi
authors.forename: Igor
authors.initials: I
authors.lastname: Del Vecchio
authors.forename: Maria Gabriella
authors.initials: MG
authors.lastname: Foti
authors.forename: Alberto
authors.initials: A
authors.lastname: Azzalin
authors.forename: Marianna
authors.initials: M
authors.lastname: Paulis
authors.forename: Elena
authors.initials: E
authors.lastname: Raimondi
authors.forename: Gabriele
authors.initials: G
authors.lastname: Vaccari
authors.forename: Umberto
authors.initials: U
authors.lastname: Agrimi
authors.forename: Giovanni
authors.initials: G
authors.lastname: Di Guardo
authors.forename: Sergio
authors.initials: S
authors.lastname: Comincini
authors.forename: Luca
authors.initials: L
authors.lastname: Ferretti
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D018076:DNA, Complementary; D023281:Genomics; D006041:Goats / Q000235:genetics*; D007438:Introns; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011110:Polymorphism, Genetic; D011328:Prions / Q000737:chemistry / Q000235:genetics*; D011401:Promoter Regions, Genetic; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D024363:Transcription Initiation Site
chemical_list: D018076:DNA, Complementary; D011328:Prions