Dept. of Pathology and Laboratory Medicine, The Children's Hospital of Philadelphia, PA 19014, USA.
Hum Immunol. 2013 Mar;74(3):325-9. doi: 10.1016/j.humimm.2012.12.007. Epub 2012 Dec 13.
Many common and well-documented (CWD) HLA alleles have only been partially characterized. The DNA sequence of these incomplete alleles, as published in the IMGT/HLA database, is most often limited to exons that code for the extracellular domains of the mature protein. Here we describe the application of next-generation sequencing technology to obtain full length genomic sequence from a single long-range PCR amplicon for 15 common and well-documented HLA Class I alleles. This technology is well suited to fill in the gaps of the current HLA allele sequence database which is largely incomplete. A more comprehensive catalog of HLA allele sequences would be beneficial in the evaluation of mismatches in transplantation, studies of population genetics, the evolution of HLAs, regulatory mechanisms and HLA expression, and issues related to the genomic organization of the MHC.
许多常见且记录良好(CWD)的 HLA 等位基因仅部分得到了描述。这些不完整等位基因的 DNA 序列,如在 IMGT/HLA 数据库中公布的,通常仅限于编码成熟蛋白胞外结构域的外显子。在这里,我们描述了应用下一代测序技术从单个长程 PCR 扩增子获得 15 个常见且记录良好的 HLA 类 I 等位基因全长基因组序列的方法。该技术非常适合填补当前 HLA 等位基因序列数据库中大部分不完整的空白。更全面的 HLA 等位基因序列目录将有助于评估移植中的错配、群体遗传学研究、HLAs 的进化、调控机制和 HLA 表达以及与 MHC 基因组组织相关的问题。