Bw.03的分子生物学研究及分子同源建模  

Molecular biological research and molecular homologous modeling of Bw.03 subgroup

作　　者：王莉[1] 孔永奎[1] 靳慧芳[1] 刘欣[1] 谢莹[1] 刘雪 常艳丽 王雅芳 杨淑淼 朱迪[1] 杨乾坤[1] WANG Li;KONG Yongkui;JIN Huifang;LIU Xin;XIE Ying;LIU Xue;CHANG Yanli;WANG Yafang;YANG Shumiao;ZHU Di;YANG Qiankun(Department of Blood Transfusion,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院输血科,河南郑州450052

出　　处：《中国输血杂志》2025年第1期112-115,共4页Chinese Journal of Blood Transfusion

摘　　要：目的对1例ABO血型B亚型进行分子生物学研究,并对突变后的酶进行三维建模预测。方法采用试管法及微柱凝胶法鉴定ABO表型;采用序列特异性引物聚合酶链反应(PCR-SSP)检测ABO血型基因,并对ABO基因第6、7外显子序列进行测序分析;通过Modeller10.4对糖基转移酶进行同源建模,应用PyMOL2.5.0软件进行分析。结果先证者标本血清学正定型B抗原减弱,反定型存在抗-B;PCR-SSP检测结果显示存在B、O基因,测序结果显示在B基因第7外显子发生c.721 C>T突变,导致p.Arg241Trp。同源建模与野生型相比,Bw.03糖基转移酶(GT)结构局部发生改变,分子间作用力分析显示241位原有的3个氢键全部消失。结论血型分子生物学检查有助于疑难血型的准确定型;同源建模能更直观的显示Bw.03 GT活性变弱的关键位点,分子间作用力分析能解释酶活性变弱的根本原因。Objective To study the molecular biological mechanism for a case of ABO blood group B subtype,and perform three-dimensional modeling of the mutant enzyme.Methods The ABO phenotype was identified by the tube method and microcolumn gel method;the ABO gene of the proband was detected by sequence-specific primer polymerase chain reaction(PCR-SSP),and the exon 6 and 7 of the ABO gene were sequenced and analyzed.Homologous modeling of Bw.03 glycosyltransferase(GT)was carried out by Modeller and analyzed by PyMOL2.5.0 software.Results The weakening B antigen was detected in the proband sample by forward typing,and anti-B antibody was detected by reverse typing.PCR-SSP detection showed B,O gene,and the sequencing results showed c.721 C>T mutation in exon 7 of the B gene,resulting in p.Arg 241 Trp.Compared with the wild type,the structure of Bw.03GT was partially changed,and the intermolecular force analysis showed that the original three hydrogen bonds at 241 position disappeared.Conclusion Blood group molecular biology examination is helpful for the accurate identification of ambiguous blood group.Homologous modeling more intuitively shows the key site for the weakening of Bw.03 GT activity.The intermolecular force analysis can explain the root cause of enzyme activity weakening.

关 键 词：Bw血型 分子生物学 同源建模 分子间作用力分析 

分 类 号：R457.11[医药卫生—治疗学] Q344.12[医药卫生—临床医学]