导致CD36缺失的基因新突变CD36c.T200A(p.Ile67Asn)  

A novel CD36 mutation c.T200A(p.Ile67Asn)that encode CD36 deficiency

作　　者：蒋丽红 吴国光 陈洁润 李丽兰 JIANG Li-hong;WU Guo-guang;CHEN Jie-run;LI Li-lan(Nanning Institute of Transfusion Medicine/Nanning Blood Center,Nanning 530003,Guangxi,CHINA)

机构地区：[1]南宁输血医学研究所南宁中心血站,广西南宁530003

出　　处：《海南医学》2022年第24期3136-3139,共4页Hainan Medical Journal

基　　金：广西自然科学基金(编号:2016GXNSFAA380143);广西南宁市科学研究与技术开发计划项目(重大科技专项)(编号:20173117);广西科学研究与技术开发计划项目(主席资金项目)(编号:08160-05)。

摘　　要：目的研究广西CD36缺失人群中发现的一个CD36基因新突变,及其导致CD36缺失的分子生物学基础。方法先证者,女性,壮族,为广西人群中检测到的一名CD36缺失个体,应用流式细胞术和血小板抗原单克隆抗体特异性免疫固定实验(MAIPA)检测确定其CD36表型;使用DNA外显子测序和c DNA克隆测序检测分析其CD36外显子和mRNA序列,针对所发现的CD36 mRNA转录本变异体,建立该变异体的真核表达细胞株,应用Western Blot验证其是否影响CD36的表达。结果先证者的CD36表型经MAIPA和流式细胞术鉴定为Ⅰ型CD36缺失。CD36 DNA外显子测序结果显示该个体具有新突变CD36c.T200A(p.Ile67Asn),为突变杂合子,CD36 c DNA克隆测序结果显示该个体有两种CD36 m RNA转录本:c.200T>A(p.Ile67Asn)(GenBank:HM217021.1)转录本变异体和CD36野生型转录本。真核表达细胞株和Western blot实验结果表明CD36转录本变异体c.200T>A可导致CD36表达缺失。结论本研究发现了一个可导致CD36缺失的基因新突变CD36 c.T200A(p.Ile67Asn),初步阐明了该个体导致CD36缺失的分子生物学基础,为CD36缺失分子机制的研究提供了依据。Objective To study a novel CD36 mutation that was found in an individual with CD36 deficiency in Guangxi,as well as its molecular basis of encoding CD36 deficiency.Methods The proband,an individual with CD36 deficiency,was female,from Zhuang ethnic group,Guangxi.The CD36 phenotype of proband was determined by flow cytometry(FCM)and monoclonal antibody immobilization of platelet antigens assay(MAIPA).CD36 exons and m RNA sequence were analyzed by DNA exons sequencing and c DNA clone sequencing to explore the molecular basis of CD36 deficiency.Eukaryotic expression cell lines were established for the CD36 m RNA transcript variant that had discovered,and its effect on CD36 expression was verified by Western Blot assay.Results CD36 phenotype of the proband was identified as type I CD36 deficiency by MAIPA and FCM.CD36 exons sequencing showed that the proband had a heterozygous mutation CD36c.T200A(p.Ile67Asn),a novel mutation in CD36 gene.CD36 c DNA cloning sequencing showed that proband has carried two kinds of CD36 mRNA transcripts:CD36 mRNA transcript variant c.200T>A(p.Ile67Asn)(GenBank:HM217021.1)and wild-type CD36 mRNA transcripts.It was verified that the CD36 m RNA transcript variant c.200T>A could result in CD36 deficiency by establishment of eukaryotic expression cell lines and Western blot assay.Conclusion This paper reports a novel CD36 mutation c.T200A(p.Ile67Asn)and preliminary elucidates the molecular basis of CD36 deficiency,providing experimental and theoretical basis for the study of the molecular mechanism of CD36 deficiency.

关 键 词：CD36缺失 基因突变 分子机制 分子克隆 细胞转染 

分 类 号：R394[医药卫生—医学遗传学]