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

Tissue Antigens. 2012 Apr;79(4):291-4. doi: 10.1111/j.1399-0039.2011.01833.x. Epub 2012 Jan 17.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles*
  • Amino Acid Motifs
  • Base Sequence
  • Crystallography, X-Ray
  • Female
  • Genomics
  • HLA-C Antigens / chemistry
  • HLA-C Antigens / genetics*
  • Humans
  • Molecular Sequence Data
  • Peptides / chemistry*
  • Protein Binding
  • Sequence Alignment

Substances

  • HLA-C Antigens
  • Peptides