title: Technical note: Advantages and limitations of authenticating Palmera goat dairy products by pyrosequencing the melanocortin 1 receptor (MC1R) gene.
abstract: Inferring the breed of origin of dairy products can be achieved through molecular analysis of genetic markers with a population-specific pattern of segregation. The goal of the current work was to generate such markers in goats by resequencing several pigmentation genes [melanocortin 1 receptor (MC1R), v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog (KIT), tyrosinase (TYR), and tyrosinase-related protein 2 (TYRP2)]. This experiment revealed 10 single nucleotide polymorphisms (SNP), including 5 missense mutations and 1 nonsense mutation. These markers were genotyped in 560 goats from 18 breeds originally from Italy, the Iberian Peninsula, the Canary Islands, and North Africa. Although the majority of SNP segregated at moderate frequencies in all populations (including 2 additional markers that were used as a source of information), we identified a c.764G>A SNP in MC1R that displayed highly divergent allelic frequencies in the Palmera breed compared with the Majorera and Tinerfeña breeds from the Canary Islands. Thus, we optimized a pyrosequencing-based technique that allowed us to estimate, very accurately, the allele frequencies of this marker in complex DNA mixtures from different individuals. Once validated, we applied this method to generating breed-specific DNA profiles that made it possible to detect fraudulent cheeses in which Palmero cheese was manufactured with milk from Majorera goats. One limitation of this approach, however, is that it cannot be used to detect illegal manufacturing where Palmero dairy products are produced by mixing milk from Palmera and Majorera goats, because the c.764G>A SNP segregates in both breeds.
authors.affiliation: Department of Animal Genetics, Center for Research in Agricultural Genomics (CSIC-IRTA-UAB-UB), Campus Universitat Autònoma de Barcelona, Bellaterra 08193, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Badaoui
authors.affiliation: Department of Animal Genetics, Center for Research in Agricultural Genomics (CSIC-IRTA-UAB-UB), Campus Universitat Autònoma de Barcelona, Bellaterra 08193, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Manunza
authors.affiliation: Department of Animal Genetics, Center for Research in Agricultural Genomics (CSIC-IRTA-UAB-UB), Campus Universitat Autònoma de Barcelona, Bellaterra 08193, Spain; Departament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Bellaterra 08193, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Castelló
authors.affiliation: Department of Agricultural, Environmental and Food Sciences, University of Molise, Via de Sanctis, 86100 Campobasso, Italy.
authors.forename: M
authors.initials: M
authors.lastname: D'Andrea
authors.affiliation: Department of Agricultural, Environmental and Food Sciences, University of Molise, Via de Sanctis, 86100 Campobasso, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Pilla
authors.affiliation: Instituto Canario de Investigaciones Agrarias, La Laguna 38108, Tenerife, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Capote
authors.affiliation: Departament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Bellaterra 08193, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Jordana
authors.affiliation: Departament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Bellaterra 08193, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Ferrando
authors.affiliation: Departamento de Genética, Universidad de Córdoba, Córdoba 14071, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Martínez
authors.affiliation: Department of Animal Genetics, Center for Research in Agricultural Genomics (CSIC-IRTA-UAB-UB), Campus Universitat Autònoma de Barcelona, Bellaterra 08193, Spain; Departament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Bellaterra 08193, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Cabrera
authors.affiliation: Departamento de Genética, Universidad de Córdoba, Córdoba 14071, Spain.
authors.forename: J V
authors.initials: JV
authors.lastname: Delgado
authors.affiliation: Departamento de Genética, Universidad de Córdoba, Córdoba 14071, Spain.
authors.forename: V
authors.initials: V
authors.lastname: Landi
authors.affiliation: Servicio de Ganadería, Diputación Foral de Bizkaia, 48014 Bilbao, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Gómez
authors.affiliation: Unitat de Races Autòctones, Servei de Millora Agrària, (SEMILLA-SAU), Son Ferriol 07198, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Pons
authors.affiliation: Livestock & Wildlife Laboratory, Arid Land Institute Medenine, 4119 Médenine, Tunisia.
authors.forename: M
authors.initials: M
authors.lastname: El Ouni
authors.affiliation: Departament de Biologia, Universitat de Girona, Girona 17071, Spain.
authors.forename: O
authors.initials: O
authors.lastname: Vidal
authors.affiliation: Department of Animal Genetics, Center for Research in Agricultural Genomics (CSIC-IRTA-UAB-UB), Campus Universitat Autònoma de Barcelona, Bellaterra 08193, Spain; Departament de Ciència Animal i dels Aliments, Universitat Autònoma de Barcelona, Bellaterra 08193, Spain. Electronic address: marcel.amills@uab.cat.
authors.forename: M
authors.initials: M
authors.lastname: Amills
mesh_terms: D000818:Animals; D004247:DNA / Q000235:genetics; D003611:Dairy Products / Q000032:analysis*; D005787:Gene Frequency; D005819:Genetic Markers; D005838:Genotype; D006041:Goats / Q000235:genetics*; D020125:Mutation, Missense; D020641:Polymorphism, Single Nucleotide; D044102:Receptor, Melanocortin, Type 1 / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D005819:Genetic Markers; D044102:Receptor, Melanocortin, Type 1; D004247:DNA
keywords: authentication of dairy products; cheese; goat; melanocortin 1 receptor gene (MC1R); pyrosequencing