转染
流式细胞术
重组DNA
分子生物学
嗜碱性粒细胞
细胞培养
免疫球蛋白E
信使核糖核酸
化学
嗜碱性粒细胞活化
生物
免疫学
生物化学
基因
抗体
遗传学
作者
Chen Chen,Qin Zhuo,Yongning Li,Hui Yang,Jingwen Zhao,Yan Li,Junsheng Huo,Xudong Jia
出处
期刊:PubMed
日期:2020-01-01
卷期号:49 (1): 80-85
标识
DOI:10.19813/j.cnki.weishengyanjiu.2020.01.014
摘要
To establish an rat basophil leukemia(RBL)-2H3 cell line stably expressing human high affinity receptor containing alpha, beta and gamma chain(hFcεRIαβγ), in order to provide experimental materials for evaluating allergenicity of food.The lentivirus was transfected into RBL-2H3 cells, and the mRNA expression of hFcεRIαβγ in cells was detected by real-time PCR and the protein expression of hFcεRIα was detected by flow cytometry.Sequencing result showed that recombinant lentiviral vector GV367-hFcεRIαβγ was successfully constructed. According to the result of experiments, lentivirus could effectively infect RBL-2H3 cells. The mRNA of hFcεRIαβγ and protein levels of hFcεRIα in RBL-2H3 cells were successfully overexpressed.The hFcεRIαβγ/RBL-2H3 cells were preliminarily constructed, which could be binded with human IgE and further used in the evaluation system of food allergy, compared to RBL-2H3 cells.
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