中国输血杂志2017，Vol.30Issue(10)：1114-1117,4.DOI:10.13303/j.cjbt.issn.1004-549x.2017.10.008
KIR3DL3基因cDNA分子克隆测序法鉴定1个常见型新等位基因
Identification of a common KIR3DL3 novel allele by molecular cDNA cloning and haplotype sequencing
摘要
Abstract
Objective To identify a novel KIR3DL3 allele in southern Chinese Han individuals by complementary DNA (cDNA) cloning and haplotype sequencing.Methods One sample with inconclusive result was identified in sequencingbased typing on the KIR3DL3 gene.We isolated the total RNA from the fresh peripheral blood of the individuals and cDNA transcripts were synthesized by RT-PCR.The KIR3DL3-specific PCR products covering the entire coding sequence of the KIR3DL3 gene was amplified by one pair of KIR3DL3-specific PCR primers,the purified PCR products were then subjected to cloning and haplotype sequencing.Results After allele separation by molecular cloning,we identified a normal KIR3DL3* 01002 allele and a novel variant allele,KIR3DL3* 04802.Nucleotide sequence alignments with KIR3DL3 alleles showed that the novel allele KIR3DL3 * 04802 differed from the closest allele KIR3DL3* 048 by a synonymous mutation at coding sequence (CDS) nt 1074 A＞G (codon337 CAA＞CAG) in exon 8.The sequence of this novel allele KIR3DL3 * 04802 was submitted to GenBank under the accession number KU529269 and the IPD-KIR Database under the submission number IWS40002178.The name KIR3DL3* 04802 has been officially assigned by the World Health Organization (WHO) Nomen clature Committee for factors of HLA system KIR subcommittee.This novel allele was identified in 12 unrelated individuals from southern Han population with a observation ratio of 3.92％.Conclusion Our results indicate that this particular assay approach utilizing cDNA cloning and the sequencing of KIR3DL3 gene presents promising applicational significance in KIR-associated studies at an allelic level.关键词
KIR3DL3基因/cDNA/分子克隆和单倍体测序/新等位基因Key words
KIR3DL3 gene/complementary DNA (cDNA)/cloning and haplotype sequencing/novel allele分类
医药卫生引用本文复制引用
邓志辉,夏华动,张国彬,陈瑞,蔡思齐..KIR3DL3基因cDNA分子克隆测序法鉴定1个常见型新等位基因[J].中国输血杂志,2017,30(10):1114-1117,4.基金项目
国家自然科学基金(81373158) （81373158）
