CAJ | 학술논문

目的：构建稳定表达人 FcεRIα( hFcεRIα)亚基的RBL-2H3细胞株。方法利用 RT-PCR技术克隆 hFcεRIα基因,构建真核表达载体 pCI-neo-hFcεRIα。脂质体介导法转染RBL-2H3细胞,G418筛选获得稳定转染细胞株。利用RT-PCR、Western blot及免疫荧光法鉴定转染结果。结果通过对脂质体介导转染体系进行优化,转染效率可高达75．38%。经 Western blot、免疫荧光及 RT-PCR 鉴定, hFcεRIα基因在RBL-2H3细胞内成功表达。结论成功构建hFcεRIα/RBL-2H3细胞模型,为深入研究IgE与FcεRI作用机制及相关药物开发提供了实验基础。
목적：구건은정표체인 FcεRIα( hFcεRIα)아기적RBL-2H3세포주。방법이용 RT-PCR기술극륭 hFcεRIα기인,구건진핵표체재체 pCI-neo-hFcεRIα。지질체개도법전염RBL-2H3세포,G418사선획득은정전염세포주。이용RT-PCR、Western blot급면역형광법감정전염결과。결과통과대지질체개도전염체계진행우화,전염효솔가고체75．38%。경 Western blot、면역형광급 RT-PCR 감정, hFcεRIα기인재RBL-2H3세포내성공표체。결론성공구건hFcεRIα/RBL-2H3세포모형,위심입연구IgE여FcεRI작용궤제급상관약물개발제공료실험기출。
Aim To construct the stable hFcεRIα/RBL-2H3 cell line expressing human FcεRIα( hFcεRIα) . Methods The human FcεRIα gene was obtained by RT-PCR and cloned into the eukaryotic expression vector pCI-neo. Then, the pCI-neo-hFcεRIα vector was transfected into RBL-2H3 cells by lipo-somes, and the transfected cells were screened through G418 fil-tration subsequently. Finally, RT-PCR, Western blot and immu-nofluorescence assay were used to determine the result of trans-fection. Results According to the optimized transfection param-eters, the transfection efficiency reached 75. 38%. The results of Western blot, immunofluorescence and RT-PCR showed that hFcεRIα could be expressed in RBL-2H3 cells successfully. Conclusion HFcεRIα/RBL-2H3 cells were successfully con-structed,which will be the experimental basis for further study on the mechanism of IgE/FcεRI and drugs for allergy diseases.