中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
CHINESE JOURNAL OF MEDICAL GENETICS
2014年
4期
511-514
,共4页
王大明%邹红岩%徐筠娉%甄建新%何柳媚%高素青
王大明%鄒紅巖%徐筠娉%甄建新%何柳媚%高素青
왕대명%추홍암%서균빙%견건신%하류미%고소청
人类白细胞抗原%新等位基因%克隆%测序
人類白細胞抗原%新等位基因%剋隆%測序
인류백세포항원%신등위기인%극륭%측서
human leackocyte antigen-C%Novel allele%Cloning%Sequencing
目的 对1例人类白细胞抗原(human leukocyte antigen,HLA)新等位基因进行序列鉴定.方法 采用QIAGEN快速DNA抽提试剂盒从外周血样本中提取基因组DNA,PCR方法扩增先证者HLA-C基因的第1外显子至第3内含子和第3内含子-3'非翻译区,PCR产物克隆转染到质粒载体中获得等位基因的单链,对所得克隆子进行第2、3和4外显子双向测序分析.结果 先证者样本克隆测序得到两个等位基因,其中一个等位基因为C* 03:04;另一个经BLAST验证为新的等位基因,序列已递交GenBank(KF049216).与最接近的C*01:03等位基因序列相比,新等位基因在第2外显子上有1个核苷酸不同,第316位C＞A,导致第82位氨基酸由精氨酸变为丝氨酸(R82S).结论 该等位基因为HLA-C新等位基因,被世界卫生组织HLA因子命名委员会正式命名为HLA-C* 01:78.先证者的HLA分型结果为A*02:07,24:02; B* 40:01,46:01; C* 01:78,03:04; DQB1* 05:02,05:02; DRB1* 16:02,16:02.
目的 對1例人類白細胞抗原(human leukocyte antigen,HLA)新等位基因進行序列鑒定.方法 採用QIAGEN快速DNA抽提試劑盒從外週血樣本中提取基因組DNA,PCR方法擴增先證者HLA-C基因的第1外顯子至第3內含子和第3內含子-3'非翻譯區,PCR產物剋隆轉染到質粒載體中穫得等位基因的單鏈,對所得剋隆子進行第2、3和4外顯子雙嚮測序分析.結果 先證者樣本剋隆測序得到兩箇等位基因,其中一箇等位基因為C* 03:04;另一箇經BLAST驗證為新的等位基因,序列已遞交GenBank(KF049216).與最接近的C*01:03等位基因序列相比,新等位基因在第2外顯子上有1箇覈苷痠不同,第316位C＞A,導緻第82位氨基痠由精氨痠變為絲氨痠(R82S).結論 該等位基因為HLA-C新等位基因,被世界衛生組織HLA因子命名委員會正式命名為HLA-C* 01:78.先證者的HLA分型結果為A*02:07,24:02; B* 40:01,46:01; C* 01:78,03:04; DQB1* 05:02,05:02; DRB1* 16:02,16:02.
목적 대1례인류백세포항원(human leukocyte antigen,HLA)신등위기인진행서렬감정.방법 채용QIAGEN쾌속DNA추제시제합종외주혈양본중제취기인조DNA,PCR방법확증선증자HLA-C기인적제1외현자지제3내함자화제3내함자-3'비번역구,PCR산물극륭전염도질립재체중획득등위기인적단련,대소득극륭자진행제2、3화4외현자쌍향측서분석.결과 선증자양본극륭측서득도량개등위기인,기중일개등위기인위C* 03:04;령일개경BLAST험증위신적등위기인,서렬이체교GenBank(KF049216).여최접근적C*01:03등위기인서렬상비,신등위기인재제2외현자상유1개핵감산불동,제316위C＞A,도치제82위안기산유정안산변위사안산(R82S).결론 해등위기인위HLA-C신등위기인,피세계위생조직HLA인자명명위원회정식명명위HLA-C* 01:78.선증자적HLA분형결과위A*02:07,24:02; B* 40:01,46:01; C* 01:78,03:04; DQB1* 05:02,05:02; DRB1* 16:02,16:02.
Objective To investigate the genetic basis for a novel allele HLA-C * 01:78.Methods Genomic DNA was extracted from peripheral blood using a QIAGEN quick DNA extraction kit.The regions encompassing HLA-C from exon 1 to intron 3 and intron 3 to 3'UTR were amplified and cloned using a cloning sequencing kit in order to split the two alleles apart.Selected clones were sequenced to include exons 2 to 4.Results Sequencing results have indicated the HLA-C alleles of the proband to be a novel C * 03:04 allele.The sequence has been submitted to GenBank (KF049216).BLAST analysis has confirmed the novel allele to have one nucleotide difference as C * 01:03 at genomic nt316C＞A (codon 82CGC＞AGC) in exon 2,which has resulted in replacement of one amino acid (82R＞S).Conclusion The novel allele has been officially named as C * 01:78 by the WHO Nomenclature Committee.The HLA allele type of the proband was therefore A* 02:07,24:02; B* 40:01,46:01; C* 01:78,03:04; DQB1 * 05:02,05:02; DRB1 * 16:02,16:02.
