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Plasmid ; 98: 31-36, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-30213734

RESUMO

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.


Assuntos
Imunoglobulina E/metabolismo , Leucemia Basofílica Aguda/metabolismo , Plasmídeos/genética , Receptores de IgE/metabolismo , Transfecção/métodos , Animais , Linhagem Celular Tumoral , Humanos , Imunoglobulina E/genética , Leucemia Basofílica Aguda/genética , Leucemia Basofílica Aguda/patologia , Ratos , Receptores de IgE/genética , Células Tumorais Cultivadas
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