摘要
目的探讨南方汉族人群KIR2DS4分子遗传多态性。方法应用特异性PCR引物扩增306名无关个体KIR2DS4基因的全部外显子，确定KIR2DS4框架基因的有无。对存在KIR2DS4的样本，对该基因的全部外显子进行双向测序，用Assign3．5分析软件鉴定基因型。在测序分型中出现与国际IPD-KIR数据库不完全匹配结果的样本，进行cDNA分子克隆和单倍型测序。结果306名南方汉族无关个体中，携带KIR2DS4基因的个体为297例，经测序分型，均获得了清晰的序列峰图。共检出了7种等位基因，其中3个新等位基因被世界卫生组织HLA因子命名委员会KIR分委会正式命名。检出的等位基因及其频率分别为KIR2DS4＊00101（78．8％）、＊003（10．5％）、＊004（16．0％）、＊010（23．2％）、＊017（0．3％）、＊00105（0．3％）和＊018（0．7％），未检出KIR2DS4＊007等位基因。能正常表达于细胞表面的等位基因（KIR2DS4＊00101、＊017、＊00105）和不能正常表达于细胞表面的等位基因（KIR2DS4＊003、＊004、＊010、＊018）的检出比例分别为79．4％及50．4％，约为1．6：1。结论获得了南方汉族人群KIR2DS4等位基因分子遗传多态性，可为移植、疾病关联研究及人类进化等提供基础数据。
Objective To study genetic polymorphisms of theKIR2DS4 gene among ethnic Hans from southern China. Methods Genomic DNA was isolated from 306 unrelated individuals and amplified with KIR2DS4 -specific PCR primers. KIR2DS4 -positive samples were genotyped for the entire coding sequence by sequencing-based typing （SBT）. Assignment of allelic genotypes was accomplished by using Assign 3.5 software. For samples with inconclusive SBT results, RT-PCR products covering the entire coding sequence of the KIR2DS4 gene were subjected to cloning and haplotype sequencing. Results Among all tested samples, 297 were demonstrated to have carried the KIR2DS4 framework gene. For KIR2DS4-positive samples subjected to SBT for the entire coding sequences, no background was observed with the obtained sequences. Three of the seven identified alleles were of novel types, which were officially named by the KIR subcommittee of the World Health Organization Nomenclature Committee for Factors of HLA System. The observed frequencies for the 7 alleles were KIR2DS4 ＊ 00101 （78.8%）, ＊003 （10.5%）, ＊ 004 （16.0%）, 010 （23.2%）, ＊ 017 （0.3%）, ＊ 00105 （0.3%） and ＊018 （0.7%）, respectively. Allele KIR2DS4 007 was not found. The overall frequency for normal cell-surface expression KIR2DS4 alleles including 2DS4 00101, ＊ 017 and ＊ 00105 was 79.4%, and that for non cell-surface expression alleles including 2DS4 003, ＊ 004, ＊ 010 and＊ 018 was 50. 4%. The ratio between the two was 1. 6 ： 1. Conclusion The present study has elucidated the allelic diversity of KIR2DS4 among ethnic Hans from southern China, which may provide valuable data for transplantation as well as studies on KIR-associated disease and evolution.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
2017年第1期21-25,共5页
Chinese Journal of Medical Genetics
基金
国家自然科学基金（81373158）
关键词
KIR2DS4框架基因
测序分型
CDNA
分子克隆
新等位基因
KIR2DS4 framework gene
Sequence-based typing
Complementary DNA
Cloning and haplotype sequencing
Novel allele
