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
Copyright policy )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
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.
Base Sequence, Amino Acid Motifs, Molecular Sequence Data, Genomics, HLA-C Antigens, Crystallography, X-Ray, Humans, Female, Peptides, Sequence Alignment, Alleles, Protein Binding
Base Sequence, Amino Acid Motifs, Molecular Sequence Data, Genomics, HLA-C Antigens, Crystallography, X-Ray, Humans, Female, Peptides, Sequence Alignment, Alleles, Protein Binding
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