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RHCE mRNA剪接体PCR检测方法的建立 被引量:4

Establishment of PCR for detecting RHCE mRNA spliceosome
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摘要 目的建立RHCE mRNA剪接体PCR检测方法,并评价方法的可行性。方法以RHCE mRNA剪接位点设计引物,提取30例随机选择的非血缘关系血液样本中的mRNA,逆转录为c DNA,并以c DNA为模板进行PCR扩增,对扩增产物进行琼脂糖凝胶电泳检测,最后通过基因测序技术对RHCE mRNA各剪接体进行序列分析。结果PCR扩增产物在2%琼脂糖凝胶中可显示出清晰的特异性条带,基因测序分析结果显示出不同剪接体的剪接位点。结论本研究建立的方法可对RHCE mRNA剪接体进行半定量检测,具有较强的可行性,为RHCE基因多态性和抗原表达的研究提供了依据。 Objective To establish the polymerase chain reaction(PCR) technique of RHCE mRNA,and to evaluate it in feasibility. Methods Primer was designed based on RHCE mRNA splicing site. The mRNA were extracted randomly from30 unrelated blood samples and were reversed transcription into c DNA and with c DNA as the template for PCR amplification.The amplified products were detected by agarose gel electrophoresis,and the RHCE mRNA splicing were sequenced by gene sequencing technique. Results PCR amplification products showed clear specific bands in 2% agarose gel and splicing sites were showed by the results of gene sequencing. Conclusion This method can be used for semi-quantitative detection of RHCE mRNA splicing. It is feasible and provides a basis for the study of RHCE gene polymorphism and antigen expression.
作者 陈鸿天 吕飘 邹敏 刘紫葳 陈婉君 刘持翔 张印则 周华友 CHEN Hongtian;LV Piao;ZOU Min;LIU Ziwei;CHEN Wanjun;LIU Chixiang;ZHANG Yinze;ZHOU Huayou
出处 《中国输血杂志》 CAS 2018年第11期1220-1222,F0001,共4页 Chinese Journal of Blood Transfusion
基金 南方医科大学南方医院人才引进科研启动经费(R10101009) 院长基金(2017C042)
关键词 RHCE MRNA 剪接体 多态性 聚合酶链反应 RHCE mRNA spliceosome polymorphism polymerase chain reaction
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  • 1邵超鹏,熊文,周一炎,安万新,刘辉,李雪梅.替代剪切形成多种形式的RhD mRNA[J].中国输血杂志,2004,17(5):310-314. 被引量:6
  • 2严力行,许先国,朱发明,何吉.RHD基因的克隆及其在K562细胞中的表达[J].中国实验血液学杂志,2005,13(3):492-495. 被引量:7
  • 3吴俊杰,许先国,洪小珍,姜侃,傅启华,严力行.一例RhD弱表达突变体的血清学和分子生物学研究[J].中华医学杂志,2005,85(23):1638-1640. 被引量:5
  • 4邵超鹏,李桢,熊文,周一炎,李雪梅.cis基因交换形成RHD-CE(2~9)-D等位基因[J].遗传,2005,27(4):561-565. 被引量:11
  • 5Avent ND, Reid ME. The Rh blood group system:a review.Blood,2000,95(2):375
  • 6Wagner FF, Flegel WA. RHCE represents the ancestral RH position, while RHD is the duplicated gene. Blood,2002,15;99(6):2272
  • 7Wagner FF, Frohmajer A, Flegel WA.RHD positive haplotypes in D negative Europeans. BMC Genetics,2001,2(1):10
  • 8Gassner C, Schmarda A, Kilga-Nogler S, et al. RHD/CE typing by polymerase chain reaction using sequence-specific primers. Transfusion,1997,37(10):1020
  • 9Rouillac C, Gane P, Cartron J, et al. Molecular basis of the altered antigenic expression of RhD in weak D(Du) and RhC/e in RN phenotypes. Blood,1996,87(11):4853
  • 10Smythe JS, Avent ND, Judson PA, et al. Expression of RHD and RHCE gene products using retroviral transduction of K562 cells establishes the molecular basis of Rh blood group antigens. Blood, 1996,87(7):2968

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