Construction and identification of recombinant RBL-2 H3 cells transfected with hFcεRIα
10.3969/j.issn.1001-1978.2015.08.029
- VernacularTitle:hFcεRIα/RBL-2 H3细胞株的构建与评价
- Author:
Nannan WANG
;
Zhongcheng LIU
;
Yanfen ZHANG
;
Yuxin LIU
;
Zhe CUI
;
Yao CHEN
- Publication Type:Journal Article
- Keywords:
hFcεRIα;
immunoglobulin E;
RBL-2H3 cell;
lipo-some transfection;
G418 filtration;
stable cell line
- From:
Chinese Pharmacological Bulletin
2015;(8):1174-1178,1179
- CountryChina
- Language:Chinese
-
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.
