TY - JOUR

T1 - Determination of cis/trans phase of variations in the MC1R gene with allele-specific PCR and single base extension

AU - Mengel-From, Jonas

AU - Børsting, Claus

AU - Sanchez, Juan J

AU - Eiberg, Hans

AU - Morling, Niels

N1 - Keywords: Alleles; Base Sequence; DNA Primers; Denmark; Electrophoresis, Capillary; Exons; Frameshift Mutation; Genetic Complementation Test; Genetic Variation; Hair Color; Humans; Phenotype; Polymerase Chain Reaction; Promoter Regions, Genetic; Receptor, Melanocortin, Type 1; Sequence Analysis, DNA; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

PY - 2008

Y1 - 2008

N2 - The MC1R gene encodes a protein with key regulatory functions in the melanin synthesis. A multiplex PCR and a multiplex single base extension protocol were established for genotyping six exonic MC1R variations highly penetrant for red hair (R), four exonic MC1R variations weakly penetrant for red hair (r), two frameshift variations highly penetrant for red hair (R) and three variations in the promoter region. We genotyped 600 individuals from Denmark using either CE or MALDI-TOF MS as the detection platform. A total of 62 individuals were genotyped R/R and among the 62 individuals, 57 had red hair and five had blond hair colour. Two different R alleles may be located in cis (RR/-) position or trans (R/R) position, and the phenotype associated with RR/- and R/R may be different. Two allele-specific PCRs were established with primers targeting the -G445A variation in the MC1R promoter and the allele-specific PCR products were used in the multiplex single base extension assay. In all 62 individuals, the MC1R variants were situated in trans position. Another 18 individuals with red hair colour were either genotyped R/- or R/r, suggesting that other genes influence hair colour.

AB - The MC1R gene encodes a protein with key regulatory functions in the melanin synthesis. A multiplex PCR and a multiplex single base extension protocol were established for genotyping six exonic MC1R variations highly penetrant for red hair (R), four exonic MC1R variations weakly penetrant for red hair (r), two frameshift variations highly penetrant for red hair (R) and three variations in the promoter region. We genotyped 600 individuals from Denmark using either CE or MALDI-TOF MS as the detection platform. A total of 62 individuals were genotyped R/R and among the 62 individuals, 57 had red hair and five had blond hair colour. Two different R alleles may be located in cis (RR/-) position or trans (R/R) position, and the phenotype associated with RR/- and R/R may be different. Two allele-specific PCRs were established with primers targeting the -G445A variation in the MC1R promoter and the allele-specific PCR products were used in the multiplex single base extension assay. In all 62 individuals, the MC1R variants were situated in trans position. Another 18 individuals with red hair colour were either genotyped R/- or R/r, suggesting that other genes influence hair colour.

U2 - 10.1002/elps.200800107

DO - 10.1002/elps.200800107

M3 - Journal article

C2 - 19016241

VL - 29

SP - 4780

EP - 4787

JO - Electrophoresis

JF - Electrophoresis

SN - 0173-0835

IS - 23

ER -