ABO基因第6外显子c.278C>T突变导致BW.12亚型的研究  

Study on BW. 12 Subtype Caused by c. 278C > T Mutation in Exon 6 of ABO Gene

作　　者：刘昕[1] 王莲慧 徐秀云[1] 舒锦[1] 乐芳 LIU Xin;WANG Lian-Hui;XU Xiu-Yun;SHU Jin;LE Fang(Department of Blood Transfusion,Jiangxi Provincial Children's Hospital,Nanchang 330006,Jiangxi Province,China)

机构地区：[1]江西省儿童医院输血科,江西南昌330006

出　　处：《中国实验血液学杂志》2022年第5期1557-1561,共5页Journal of Experimental Hematology

基　　金：江西省卫生健康委科技计划(SKJP220201919)。

摘　　要：目的:研究ABO基因α-1,3-D半乳糖基转移酶基因突变对B抗原表达的影响,探讨其分子机制。方法:利用常规血清学方法对先证者及其家系成员进行血型鉴定,应用聚合酶链反应-序列特异性(PCR-SSP)和ABO基因第1-7外显子PCR产物直接测序进行ABO基因分型及序列分析,应用生物信息学软件对突变蛋白进行3D结构模拟,分析基因突变对蛋白结构稳定性的影响。结果:先证者及其家系成员血清学检测结果为B亚型,ABO基因分型明确了先证者的基因型为Bw12/O;基因测序结果证实B等位基因第6外显子存在ABO*BW. 12的特征性变异c. 278C> T,导致多肽链p. Pro93Leu替换。蛋白3D结构的模拟分析发现氨基酸替换后,与93位氨基酸连接的氢键及水分子均发生了改变。家系调查发现先证者的爷爷、父亲、大伯和哥哥均携带了相同的ABO*BW. 12等位基因。结论:ABO基因第6外显子c. 278C> T突变导致多肽链氨基酸替换,影响了α-1,3-D半乳糖基转移酶蛋白稳定性,引起酶活性改变,产生Bw12表型,且该变异基因可稳定遗传。Objective:To investigate the effect of ABO geneα-1,3-D galactosyl transferase mutation on B antigen expression and its molecular mechanism. Methods:The proband and their family members were identified by routine serological methods,and ABO genotyping and sequence analysis were performed by polymerase chain reaction-sequence specificity(PCR-SSP)and direct sequencing of PCR products from exon1-7of ABO gene. The3D structural simulation of mutant proteins was performed by bioinformatics software. The effect of gene mutation on protein structural stability was analyzed. Results:The proband and his family members were subtype B. ABO genotyping indicated that the proband′s genotype was Bw12/O. Gene sequencing results confirmed the presence of ABO*BW. 12 characteristic variation c.278C>T in the6th exon of allele B,leading to the replacement of polypeptide chain p. Pro93Leu. The3D structure simulation analysis of the protein showed that the hydrogen bonds and water molecules connected to the protein changed after amino acid substitution. The family investigation found that the grandfather,father,uncle and brother of the proband all carried the same ABO*BW. 12 allele. Conclusion:The mutation of the6th exon c.278C>T of ABO gene led to the substitution of polypeptide chain amino acids,which affected the stability ofα-1, 3-D galactosyl transferase protein,resulting in the change of enzyme activity,and the Bw. 12 phenotype,which can be stably inherited.

关 键 词：ABO基因 基因分型 基因测序 3D结构模拟 

分 类 号：R457.11[医药卫生—治疗学]