中国输血杂志2017，Vol.30Issue(10)：1106-1109,4.DOI:10.13303/j.cjbt.issn.1004-549x.2017.10.006
KIR2DL4基因测序分型中杂合碱基位置峰高不平衡现象及其意义
The in-balance peak height at heterozygous nucleotide positions observed in KIR2DL4 sequencing-based typing and its significance
摘要
Abstract
Objective To explore the reason for unusual sequences and genotyping results in sequencing-based typing (SBT) for KIR 2DL4 gene.Methods Two samples with in-balance peak height at heterozygous nucleotide positions were observed during KIR 2DL4 SBT,in which one sample was assigned with ambiguous SBT result and the other one was ob served with no conclusive SBT results using the Assign 4.7 software.We isolated total RNA from fresh peripheral blood and complementary DNA (cDNA) transcripts was synthesized by RT-PCR,and then subjected to cloning and haplotype sequencing.The copy number of KR2DL4 was determined by quantitative PCR reactions.Results The results of molecular cloning and haplotype sequencing demonstrated that one sample with three normal alleles KIR 2DL4* 00102,* 00501,* 011;and the other sample with three normal KIR 2DL4 * 00102,* 00501 and * 00602 alleles were determined.No novel KIR 2DL4 al lele was identified.Both the two samples carry a copy number of 3 KIR 2DL4 genes determined by quantitative PCR assay.Conclusion KIR 2DL4 gene can carry extra diversity of copy number variation in human KIR haplotype,samples with in balance peak height at heterozygous nucleotide positions and no conclusive result in KIR 2DL4 SBT test can't be simply explained by putative novel alleles.关键词
KIR 2DL4基因/测序分型/基因拷贝/杂合碱基/峰高Key words
KIR 2DL4 gene/sequencing-based typing/copy number/heterozygous nucleotide/peak height分类
医药卫生引用本文复制引用
喻琼,甄建新,张国彬,陈瑞,蔡思齐,邓志辉..KIR2DL4基因测序分型中杂合碱基位置峰高不平衡现象及其意义[J].中国输血杂志,2017,30(10):1106-1109,4.基金项目
国家自然科学基金(81373158) （81373158）
