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

发布时间：2018-02-24 16:30

本文关键词： HLA抗原等位基因组织工程干细胞移植人类白细胞抗原E PCR-SBT 全基因序列测定新等位基因多态性　出处：《中国组织工程研究》2017年25期 　论文类型：期刊论文

【摘要】：背景:人类白细胞抗原E是非经典的人类白细胞抗原I类分子,目前的多态性分析研究主要针对其第3外显子人类白细胞抗原E~*01:01及人类白细胞抗原E*01:03进行区分,然而其全基因序列测定方法及新等位基因报道尚不多见。目的:利用中国深圳地区无偿献血正常个体建立人类白细胞抗原E基因全长序列测序分型方法,鉴定人类白细胞抗原E基因全长序列新的等位基因。方法:提取研究对象外周血DNA,根据IMGT/HLA数据库中公布的人类白细胞抗原E基因序列,在保守区设计全基因扩增及测序引物,利用高保真反应体系对人类白细胞抗原E全长序列进行扩增,随后测序、拼接、序列确认及基因定型。结果与结论:(1)成功建立了人类白细胞抗原E的全基因扩增及测序分型方法,发现了两个人类白细胞抗原E新等位基因——人类白细胞抗原E~*01:01:01:06与人类白细胞抗原E~*01:01:01:07,获得了WHO人类白细胞抗原命名因子委员会的命名;(2)人类白细胞抗原E~*01:01:01:06与其最接近的等位基因人类白细胞抗原E~*01:01:01:01相比,突变位点在5’-启动子区的NT-26(G-T),人类白细胞抗原E~*01:01:01:07与其最接近的等位基因人类白细胞抗原E~*01:01:01:01相比,突变位点在3’-非翻译区的NT3345位(T-C);(3)结果提示,中国人群中人类白细胞抗原E全基因序列多态性数据有待填补,试验建立的方法为该项研究提供了关键的技术。
[Abstract]:Background: human leukocyte antigen E is a nonclassical class I molecule of human leukocyte antigen. The present polymorphism analysis is mainly focused on the differentiation of human leukocyte antigen Et01: 01: 01 and human leukocyte antigen Et01: 03 at exon 3. However, the whole gene sequencing method and the new alleles have not been reported. Objective: to establish a full-length sequence sequencing method for human leukocyte antigen E gene from normal volunteers in Shenzhen area, China. Methods: a novel allele of human leukocyte antigen E gene was identified. Methods: the whole gene amplification and sequencing primers were designed according to the sequence of human leukocyte antigen E gene published in IMGT/HLA database. The full-length sequence of human leukocyte antigen E was amplified by high fidelity reaction system, then sequenced and spliced. Results and conclusion the whole gene amplification and sequencing of human leukocyte antigen E were successfully established. Two new alleles of human leucocyte antigen E were found: human leukocyte antigen E 01: 01: 01: 06 and human white blood cell antigen 01: 01: 01: 07. The name of the WHO Committee of Human Leukocyte Antigen nomenclature factors was obtained. The nearest allele, the human leukocyte antigen, was compared to 01: 01: 01: 01. The mutation locus is in the NT-26G-TN of the 5NT-promoter region, and the NT3345 position T-C3 of the human leukocyte antigen Et01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01: 01. The data of human leukocyte antigen E gene polymorphisms in Chinese population need to be filled. The experimental method provides a key technique for this study.
【作者单位】：深圳市血液中心深圳市组织配型与免疫遗传重点实验室;
【基金】：广东省医学科研基金(A2016222)资助深圳市科技研发资金(GJHZ20160229170608241)~~
【分类号】：R440

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