人ABO基因5’非翻译区的基因克隆和序列分析  

Cloning and sequence analysis of 5＇-untranslated region of human ABO gene

作　　者：刘瑛[1] 许先国[1] 马开荣[1] 兰小飞[1] 洪小珍[1] 朱发明[1] 吕杭军[1] 严力行[1] 

机构地区：[1]浙江省血液中心输血研究所,卫生部血液安全研究重点实验室,杭州310006

出　　处：《中华医学遗传学杂志》2011年第1期83-87,共5页Chinese Journal of Medical Genetics

基　　金：国家自然科学基金（30871112）;浙江省医药卫生优秀青年科技人才基金（2008QN008）;浙江省卫生高层次创新人才基金

摘　　要：目的建立ABO基因5’非翻译区（5’untranslated region，5’-UTR）长片段的序列分析方法，研究5CUTR的基因多态性，为揭示AB0基因上游转录调控的分子机制提供基础。方法以血型血清学方法和分子生物学方法鉴定30名献血者的ABO表型和第6和7外显子基因型，采用PCR长片段扩增技术得到ABO基因5＇-UTR约5kb产物，直接进行双向测序分析，并通过克隆对ABO基因5＇-UTR存在杂合的样本进行单倍型分析。结果在表型和基因型相符合的样本中，直接测序法在5＇-UTR共发现20个多态性位点，包括16个单核苷酸多态性位点、1个6bp重复序列插入／缺失多态性、1个8bp重复序列缺失多态性、1个36bp插入多态性、1个43bp重复序列多态性，其中11个为新发现的多态性位点。单倍型分析确定这些变异位点的顺／反式连锁关系，发现ABO基因5＇-UTR的7种单倍型序列组合。结论人ABO基因5＇-UTR具有序列多态性，启动子近端的序列相对保守。Objective To clone and investigate the polymorphism of the 5＇ untranslated region （5＇ UTR） of the human ABO gene, in order to provide the basis for exploring the transcriptional regulation of the human ABO histo-blood group genes. Methods ABO phenotypes of 30 unrelated healthy blood donors were determined by serological technique, their genotypes were analyzed by sequencing the exons 6 and 7 of the ABO gene. Nearly 5 kb of the 5＇-UTR of ABO gene was obtained by PCR amplification and sequencing was performed bidirectionally. Haplotypes of samples with heterozygous sites in the 5CUTR of ABO gene were separated and analyzed after cloning. Results Twenty polymorphic sites were identified in these samples where ABO genotypes were consistent with serological phenotypes. It included sixteen nucleotide sequence variations, one 8 bp deletion, one 6 bp deletion/insertion, one 36 bp-insertion and one 43 bp repeats. Among them, 11 were novel polymorphic sites. Seven different haplotypes of 5＇-UTR of ABO gene were defined to the cis/trans linkage of those mutations. Conclusion There were polymorphisms in the 5C UTR of ABO gene and the nucleotide sequences near the proximal promoter were conservative.

关 键 词：5’非翻译区 ABO血型 基因型 单倍型 多态性 

分 类 号：R394[医药卫生—医学遗传学]