中国药理学通报Issue(8)：1174-1178,1179,6.DOI:10.3969/j.issn.1001-1978.2015.08.029
hFcεRIα/RBL-2 H3细胞株的构建与评价
Construction and identification of recombinant RBL-2 H3 cells transfected with hFcεRIα
摘要
Abstract
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.关键词
hFcεRIα/IgE/RBL-2H3 细胞/脂质体转染/G418 筛选/稳定细胞株Key words
hFcεRIα/immunoglobulin E/RBL-2H3 cell/lipo-some transfection/G418 filtration/stable cell line分类
医药卫生引用本文复制引用
王南南,刘中成,张艳芬,刘玉欣,崔哲,陈瑶..hFcεRIα/RBL-2 H3细胞株的构建与评价[J].中国药理学通报,2015,(8):1174-1178,1179,6.基金项目
国家自然科学基金资助项目( No 81202338) （ No 81202338）
河北省自然科学基金资助项目( No H2013201128) （ No H2013201128）
中国博士后科学基金资助项目(No 2013M530885) （No 2013M530885）
河北省教育厅项目(No Z2015013) （No Z2015013）
