新等位基因c.175_(1)76insGA致B_(el)亚型的家系分析  

Pedigree analysis of B_(el) subtype caused by the new allele c.175_176insGA

作　　者：杨贺才[1] 关茵[2] 马晓莉[1] 吕永磊[1] 孔永奎[3] 郭超群[2] 耿明璐[1] 王丽萍 温涛[1] Yang Hecai;Guan Yin;Ma Xiaoli;Lyu Yonglei;Kong Yongkui;Guo Chaoqun;Geng Minglu;Wang Liping;Wen Tao(Laboratory of Blood Group Research,Henan Red Cross Blood Center,Zhengzhou 450000,China;Quality Control Department,Jiaozuo Central Blood Station,Jiaozuo 454001,China;Department of Blood Transfusion,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]河南省红十字血液中心血型研究室,郑州450000 [2]焦作市中心血站质控科,焦作454001 [3]郑州大学第一附属医院输血科,郑州450052

出　　处：《中华检验医学杂志》2024年第10期1206-1211,共6页Chinese Journal of Laboratory Medicine

基　　金：河南省医学科技攻关计划(LHGJ20220260,RKX202101005)。

摘　　要：目的对1例B_(el)亚型新等位基因c.175_(1)76insGA的家系进行血清学和基因型分析,并初步探讨糖基转移酶B弱表达的分子机制。方法描述性研究,选取1例23岁的男性无偿献血者及其家系成员为研究对象。采用试管法鉴定先证者及其家系成员的ABO和Le血型,应用凝集抑制试验检测唾液中B和H抗原,并对研究对象进行Sanger测序和PacBio(Pacific Bioscience)三代技术单倍型测序,鉴定基因型。最后应用Expasy软件对DNA序列进行氨基酸翻译并预测基因改变后的蛋白长度,应用ORF finder预测替代起始密码子以及mRNA开放阅读框,并进行蛋白表达机制分析。结果先证者及其姐姐血清学为B_(el)亚型,其母亲为AB_(el)亚型,其父亲为正常O型,家系成员均为Le(a+b+)表型。Sanger测序结果表明,在先证者及其母亲和姐姐第4外显子均发现c.175_(1)76insGA新等位基因。三代单倍型测序检测家系成员的单倍型发现,先证者ABO*O.01.02/ABO*BEL.NEW(c.175_(1)76insGA),父亲ABO*O.01.02/ABO*O.01.02,母亲ABO*A1.02/ABO*BEL.NEW(c.175_(1)76insGA),姐姐ABO*O.01.02/ABO*BEL.NEW(c.175_(1)76insGA)。蛋白表达机制分析表明,虽然据推测ABO*BEL.NEW新等位基因发生了移码突变,在第5外显子提前出现了终止密码子p.Asp59Glu*fs20,编码无活性的糖基转移酶。但可利用替代起始密码子启动功能性B_(el)亚型糖基转移酶的翻译。结论B_(el)亚型新等位基因的表达与替代起始密码子启动B_(el)亚型糖基转移酶的翻译有关。ObjectiveTo serologically and genotypically analyze the pedigree of a case with a new allele c.175_176insGA of B_(el)subtype and preliminarily explore the molecular mechanism of weak expression of glycosyltransferase B.MethodIn the descriptive study,a 23-year-old male voluntary blood donor and his family members were selected for the study.The ABO and Le blood types of the proband and his family members was identified by the test tube method.The agglutination inhibition test was applied to detect the B and H antigens in saliva,and the Sanger sequencing and PacBio(Pacific Bioscience)third-generation haplotype sequencing were performed on the study subjects to identify genotypes.Finally,Expasy software were applied to amino acid translation of DNA sequences and prediction of protein length after gene alteration.ORF finder was applied to predict alternative start codons as well as open reading frames of mRNA,and protein expression mechanisms were analyzed.ResultsThe proband and her sister were B_(el)subtype,her mother was AB_(el)subtype,her father was normal O type,and all members of the family were Le(a+b+)phenotype.Sanger sequencing results showed that a new allele of c.175_176insGA was found in exon 4 of the proband,her mother,and her sister.Third-generation haplotype sequencing detected the haplotypes of the family members,which revealed that the proband was ABO*O.01.02/ABO*BEL.NEW(c.175_176insGA),the father was ABO*O.01.02/ABO*O.01.02,the mother was ABO*A1.02/ABO*BEL.NEW(c.175_176insGA),and the sister was ABO*O.01.02/ABO*BEL.NEW(c.175_176insGA).Analysis of the protein expression mechanism indicated that although the new allele of ABO*BEL.NEW was presumed to cause a frameshift mutation and result in a premature stop codon p.Asp59Glu*fs20 in exon 5,encoding an inactive glycosyltransferase,an alternative start codon could be utilized to initiate translation of B_(el)subtype functional glycosyltransferase.ConclusionExpression of the new allele of B_(el)subtype is associated with the translation of B_(el)subtype gly

关 键 词：B_(el)亚型 基因型 糖基转移酶 蛋白表达 

分 类 号：R457.11[医药卫生—治疗学]