ABO血型新等位基因Ael08的分子生物学研究  

Molecular basis of a novel allele Ael08

作　　者：吕杭军[1] 洪小珍[1] 应燕玲[1] 许先国[1] 朱发明[1] 严力行[1] 

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

出　　处：《中国输血杂志》2011年第8期676-679,共4页Chinese Journal of Blood Transfusion

基　　金：国家自然科学基金(30871112);浙江省医药卫生科研基金(2011RCB012)

摘　　要：目的研究ABO新等位基因Ael08的分子机制。方法利用单克隆抗体检测先证者标本红细胞ABO血型抗原，标准A、B、O红细胞检测血清中的ABO抗体，吸收放散试验检测红细胞上微量抗原；采用PCR技术扩增先证者ABO基因的第5～7外显子序列，PCR产物经双酶切后直接测序分析6和7外显子；扩增产物经TOPOTA克隆到质粒载体中获得单链，对所得克隆作ABO基因第6、7外显子双向测序分析；家系调查采集先证者父亲和哥哥的标本作血型血清学试验和ABO基因第6和7外显子直接测序分析。结果先证者红细胞上表达很弱的A抗原，只有通过吸收放散才能有效检出，同时其血浆中存在较强的抗-B和较弱的抗-A；直接测序分析发现：6号外显子第261位缺失杂合，7号外显子第297位A／G、467C／T、646T／A、681G／A、771C／T、829G／A、804insG／G杂合；克隆测序发现1个为常见的002等位基因，另1个为1种新的等位基因（已被dRBCNCBI命名为Ae抑8），与A102比较，Ael08在第804位碱基处插入G，这导致氨基酸第268位后阅读框架发生改变，编码产物比正常A1转移酶多37个氨基酸。家系调查显示：先证者Ael08等位基因从父亲遗传所得。结论发现1个ABO新等位基因Ael08，α-1，3-N．乙酰半乳糖胺基转移酶基因（A基因）第804位碱基处插入G突变导致产生Ael表型。Objective To analyze the molecular genetic basis for Ael08 allele of ABO blood group. Methods The ABO group antigens on red blood cells of the proband were identified by monoclonal antibodies and the ABO antibody in se- rum was detected by the standard A, B, O cells. The weak antigen on RBCs was detected by absorption-elution test. Exon 5 to exon 7 encoding region ofABO gene was amplified by polymerase chain reaction （PCR） and the PCR product was se- quenced directly for exon 6 and 7 after double enzyme digestion. The amplified product for exon 5 to 7 was also cloned by TOPO TA cloning sequencing kit to split the two alleles apart and the chosen colonies were＇sequenced bidirectionally for ex- on 5 to 7 of ABO gene. The samples of the father and brother of the proband were collected, then the blood group serological test and sequence analysis for exon 6 to 7 of ABO gene were performed. Results Weak A antigen was found on the red cells of the proband,identified only by absorption-elution test and there was and-B and weak anti-A antibody in his serum. There was 261G deletion heterozygote and 297A/G in exon 6 and 467C/T,646T/A,681G/A,771C/T,829G/A,804insG/G heterozygotes in exon 7 by directly DNA sequencing. After claning and sequencing, one common allele 002 and a novel ABO allele were obtained, the latter was nominated as Ael08 by dRBC NCBI. The sequence of Ael08 has one nucleotide change at 804 position G insertion compared with that orAl02 ,which results in reading frameshifi and encoded a product 37 amino acids longer than that of A1 transferase. The Ael08 in the proband was inherited from his father by family investigation. Conclusion A novel allele Ael08 was identified and insertion G at position 804 of N-acetyl-galactos-aminyltrans- ferase gene （A gene） can result in Ael phenotype.

关 键 词：Ael表型 ABO基因 抗-A 新等位基因 

分 类 号：R457.11[医药卫生—治疗学]