MNS血型相关基因GYPA、GYPB、GYPE mRNA全长序列多态性分析  

Polymorphism of the Full-Length mRNA Sequences of MNS blood group-related genes GYPA,GYPB and GYPE

作　　者：梁延连[1] 吴凡[1] 梁爽 徐筠娉[1] 苏宇清[1] LIANG Yan-Lian;WU Fan;LIANG Shuang;XU Yun-Pin;SU Yu-Qing(Institute of Blood Transfusion Medicine,Shenzhen Blood Center,Shenzhen 518000,Guangdong Province,China)

机构地区：[1]深圳市血液中心输血医学研究所,广东深圳518000

出　　处：《中国实验血液学杂志》2023年第5期1537-1542,共6页Journal of Experimental Hematology

基　　金：广东省医学科研基金(B2020179);广东省自然科学基金(2020A1515010387);深圳市医疗卫生三名工程(SZSM2O1811092);深圳市输血医学重点学科(SZXK070)。

摘　　要：目的:分析MNS血型相关基因GYPA、GYPB、GYPE mRNA全长序列的特征,了解MNS血型基因的多态性。方法:随机选取无偿献血者500人份离体24 h内的抗凝血各8 ml,以血型血清学方法鉴定MN、Ss、Mia血型。从500例样本中随机选取MNS与Mia血型不同组合的5例样本,使用密度梯度离心法分离外周血中的单个核细胞(PBMC)并提取总mRNA,使用逆转录试剂盒制备cDNA,采用巢式PCR(nested PCR)方法扩增目标片段,切胶回收后分别对GYPA、GYPB、GYPE mRNA全长序列进行测序,通过Oligo 6.0软件分析碱基序列。结果:血清学检测得到MN、Ss和Mia表现型,不同个体间的红细胞与抗-Mia血清反应存在凝集强度差异。检测了5例不同表现型组合样本的GYPA、GYPB、GYPE基因mRNA全长序列,1例样本的GYPA mRNA中exon-6完整缺失,其他4例样本的GYPA mRNA全长完整;2例样本的GYPB mRNA中exon-2完整缺失,Mia血型阴性;2例样本的GYPB mRNA全长完整,Mia血型阳性;1例样本的GYPB mRNA中exon-5存在碱基替换;5例样本的GYPE mRNA全长完整。结论:MNS血型相关基因具有明显的多态性,mRNA全长序列的检测为GYPA、GYPB、GYPE基因结构的分析与MNS血型抗原表达的深入研究奠定了基础。Objective:The characteristics of the full-length mRNA sequences of MNS blood group-related genes GYPA,GYPB and GYPE were analyzed to understand the polymorphism of MNS blood group genes.Methods:Anticoagulated blood within 24 h from 500 unpaid blood donors(8 ml each)were randomly selected,and MN,Ss and Mia blood types were identified by serological methods.5 samples with different combinations of MNS and Mia blood types were randomly selected from 500 samples,and peripheral blood mononuclear cells(PBMC)were isolated by density gradient centrifugation,then total mRNA was extracted.cDNA was prepared by using the reverse transcription kit.The target fragments were amplified by nested PCR,and the full-length mRNA sequences of GYPA,GYPB and GYPE were sequenced after gel cutting and recycling,and the base sequences were analyzed by Oligo 6.0 sof tware.Results:The MN,Ss and Mia phenotypes were detected by serological methods,and there were differences in agglutination intensity of red blood cells(RBC)and anti-Mia serum between different individuals.The full-length mRNA sequences of GYPA,GYPB and GYPE genes in 5 samples of different phenotype combinations were detected.The exon-6 was completely deleted from the GYPA mRNA in 1 sample,and the full-length of GYPA mRNA in the other 4 samples were complete.The exon-2 was completely deleted from the GYPB mRNA in 2 samples,with Mia blood type negative.2 samples showed complete full-length of GYPB mRNA,with Mia blood type positive.There was base substitution in exon-5 of GYPB mRNA in 1 sample.The full-length of GYPE mRNA was intact in 5 samples.Conclusion:MNS blood group related-genes have obvious polymorphism,and the detection of full-length mRNA sequence lays a foundation for the analysis of GYPA,GYPB and GYPE gene structure and in-depth study of MNS blood group antigen expression.

关 键 词：MNS血型 mRNA全长序列 GYPA、GYPB、GYPE基因 选择性RNA剪接 

分 类 号：R446.6[医药卫生—诊断学]