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)