转染
脂质体
分子生物学
细胞培养
生物
重组DNA
污渍
绿色荧光蛋白
质粒
载体(分子生物学)
细胞
U937电池
免疫球蛋白E
基因
免疫学
抗体
生物化学
遗传学
作者
Zhongcheng Liu,Lifang Hao,Nanan Wang,Su Zhang,Nan Zhang,Zhenzhen Xu,Yanlei Yang,Yanfen Zhang
出处
期刊:Plasmid
[Elsevier]
日期:2018-06-01
卷期号:98: 31-36
被引量:1
标识
DOI:10.1016/j.plasmid.2018.09.004
摘要
IgE/FcεRI signal pathway plays a crucial role in triggering allergic reactions, and there is no cross-recognition between IgE and FcεRI in human and rats. In order to obtain the hFcεRIα/ RBL-2H3 cell line, total RNA was extracted from U937 cells, and the human FcεRIα gene was obtained by RT-PCR technology. Then the amplified product was digested and inserted into the pIRES2-EGFP vector. After the plasmid was transfected into the RBL-2H3 cells using lipofectamine, and the RBL-2H3 cell lines of stable expression were screened by G418. The transfection efficiency reached 60.45% with optimizing transfection parameters. The last the expression of hFcεRIα was detected by RT-PCR, western blotting and fluorescent microscopy. The present results demonstrated that the pIRES2-EGFP-hFcεRIα vector was constructed and a stable cell line of hFcεRIα/ RBL-2H3 cells was established successfully. This cell line is promising tools for further research on the pathogenesis and drug development of allergic diseases.
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