title: Allele-specific amplification of the complete HLA-C gene from genomic DNA - a novel Cw4 allele (C*04:71) with a Cw1 motif in the peptide-binding site.
abstract: To determine the complete sequence of a newly identified human leukocyte antigen (HLA)-C allele, we designed a method where the full genomic sequence of HLA-C*04 was amplified in isolation from the patient second HLA-C allele in a single polymerase chain reaction (PCR), using primers spanning its 5'- and 3'-untranslated regions. The new allele, officially designated HLA-C*04:71, differs from HLA-C*04:01:01:01 by two single-nucleotide polymorphisms: one determines substitution of phenylalanine for serine 9 at the B pocket of the peptide-binding site; the second substitution is a new polymorphism in intron 5. Phe-9 is characteristic of Cw1 alleles and its presence in C*04:71 most likely affects its peptide-binding repertoire. The principle we have used for C*04:71 isolation could be adapted for unambiguous sequence-based HLA-C typing of selected samples in a clinical setting.
authors.affiliation: Laboratorio de Inmunogenética-HLA, Servicio de Inmunología, Hospital Universitario Puerta de Hierro, Majadahonda, Madrid, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Moraru
authors.forename: A
authors.initials: A
authors.lastname: Balas
authors.forename: R
authors.initials: R
authors.lastname: de Pablo
authors.forename: J L
authors.initials: JL
authors.lastname: Vicario
authors.forename: C
authors.initials: C
authors.lastname: Vilches
mesh_terms: D000483:Alleles; D020816:Amino Acid Motifs; D001483:Base Sequence; D018360:Crystallography, X-Ray; D005260:Female; D023281:Genomics; D015236:HLA-C Antigens / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D016415:Sequence Alignment
chemical_list: D015236:HLA-C Antigens; D010455:Peptides