Identification of genomic full-length sequence of human leukocyte antigen-E and its two novel alleles
10.3969/j.issn.2095-4344.2017.25.022
- VernacularTitle:人类白细胞抗原E基因全长序列测定及两个新等位基因的鉴定
- Author:
Liumei HE
;
Songxing WANG
;
Yunping XU
- From:
Chinese Journal of Tissue Engineering Research
2017;21(25):4068-4074
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Human leukocyte antigen-E (HLA-E) is one of non-classical HLA class I genes. Up to now, the polymorphism analysis is mainly aimed at the variation in exon 3 of HLA-E, which determines HLA-E*01:01 or HLA-E*01:03. However, the identification of the full-length HLA-E and its novel alleles is rare reported.OBJECTIVE: To establish the method of identification of HLA-E genomic full-length sequence, and to identify its novel alleles in healthy blood donors in Shenzhen, China.METHODS: Peripheral blood DNA samples were extracted from the subjects, and the amplified primers and sequencing primers in conserved regions were designed according to the sequences of HLA-E published in the IMGT/HLA database.A high-fidelity reaction system was used to amplify the genomic full-length of HLA-E, followed by sequencing,assembling, confirming and typing.RESULTS AND CONCLUSION: Herein, we successfully established the method for amplifying genomic full-length sequence and sequence-based typing. Two novel HLA-E alleles were nominated by WHO HLA Nomenclature committee as HLA-E*01:01:01:06 and HLA-E*01:01:01:07. Compared with the most related allele HLA-E*01:01:01:01,HLA-E*01:01:01:06 had one nucleotide change at nt-26(G->T) in 5'-promoter, and HLA-E*01:01:01:07 had one nucleotide change at nt3345(T->C) in 3'-UTR. The polymorphism data of genomic full-length HLA-E in Chinese individuals need to be filled, and the method we developed here supplies the key technique for the further studies.
