关键词: 人类白细胞抗原, 基因组序列, 克隆测序, 调控区, 多态性

Abstract: In the present study, a high-resolute method for cloning and sequencing genomic full-length HLA-A and -B using 20 Chinese Han individuals was established. We detected 10 HLA-A allele sequences 4.2 kb in length and 6 HLA-B allele sequences 3.7 kb in length, and the sequences included all exons, all introns, 5′promoter, and 3′UTR of the two genes. All sixteen sequences have been submitted to GenBank and IMGT/HLA database. A*1153 is a novel allele, and the introns of B*151101 are firstly reported here. The 5′promoter and 3′UTR sequences of 5 HLA-A alleles and 2 HLA-B alleles are also firstly disclosed, and all other alleles have extended the genomic full length sequences released in IMGT/HLA database. The polymorphic structures of upper 5′promoter and downstream 3′UTR, which were uncovered in IMGT/HLA database, are firstly depicted in Chinese Han individuals. Twenty-six single nucleotide polymorphisms (SNPs) and one 3 bp-insertion/deletion (Indel) were located in the upper 5′promoter and 14 SNPs were located in the 3′UTR of HLA-A. In addition, five SNPs and one 1 bp-indel were located in the upper 5′promoter and 5 SNPs were located in the 3′UTR of HLA-B. Through analyzing the phylogenetic trees of 5′promoter, exons and 3′UTR of the two genes, we found that the evolution history of regulatory regions and exons is different between the two genes. The regulatory regions are tightly linked with exons in most of HLA-A alleles excluding A*24020101. On the contrary, recombinant events may occur fre-quently between regulatory regions and exons in most HLA-B alleles.

Key words: polymorphism, human leucocyte antigen (HLA), genomic sequence, cloning and sequencing, regulatory region