Abstract
Despite several studies that defined the polymorphism of the nonclassical human leukocyte antigen-E (HLA-E), HLA-F, and HLA-G genes, most polymorphisms thus far examined in correlative studies were derived from the coding sequences of these genes. In addition, some discrepancies and ambiguities in the available data have persisted in current databases. To expand the data available and to resolve some of the discrepant data, we have defined protocols that allow for the amplification of 6 to 7 kb of contiguous genomic sequence for each gene, including all of the coding and intron sequences, approximately 2 kb of 5' flanking promoter sequence, and 1 kb of 3' flanking sequence. Using long-range polymerase chain reaction (PCR) protocols, generating either one or two PCR products depending on the locus, amplified genomic DNA was directly sequenced to completion using a set of about 30 primers over each locus to yield contiguous sequence data from both strands. Using this approach, we sequenced 33 genomic DNAs, from Asian, African American, and Caucasian samples. The results of this analysis confirmed several previously reported coding sequence variants, identified several new allelic variants, and also defined extensive variation in intron and flanking sequences. It was possible to construct haplotype maps and to identify tagging single nucleotide polymorphisms that can be used to detect the composite variation spanning all three genes.
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Acknowledgements
The expert technical assistance of Scott Medley is gratefully acknowledged. This work was supported by National Institute of Health grants AI33484 and AI49245 to DEG. A.I. was supported in part by Grant in Aid number 05671391 from the Ministry of Education, Science, and Culture in Japan and by National Institutes of Health grant AI49213. L.P.Z. was supported by CA106320.
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Pyo, CW., Williams, L.M., Moore, Y. et al. HLA-E, HLA-F, and HLA-G polymorphism: genomic sequence defines haplotype structure and variation spanning the nonclassical class I genes. Immunogenetics 58, 241–251 (2006). https://doi.org/10.1007/s00251-005-0076-z
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DOI: https://doi.org/10.1007/s00251-005-0076-z