[Establishment of C26 cell strain stably expressing folate receptor α] (original) (raw)

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology, 2015

Abstract

Objective To establish a C26 cell strain stably expressing folate receptor α (FRα) for the subsequent study of FRα DNA vaccine. Methods C26 cells were transfected with previously constructed recombinant eukaryotic expressing vector pcDNA3.1-FRα by Lipofectamine(TM)2000. Afterwards the cells were subjected to G418 (500 mg/mL) selection to get G418 resistant cells. And then single cell cloning was performed to generate monoclonal cell strain. The gene and protein expression levels of FRα of the monoclonal cell strain were further analyzed by reverse transcription PCR (RT-PCR) and fluorescence microscopy. The cells were cultured for several generations, and the mRNA expression of FRα was analyzed by RT-PCR at different generations to determine whether the transfected gene was stable or not during cell passaging. Results After transfection, G418 selection and single cell cloning, the monoclonal cell strain were established and proved to be able to express FRα mRNA and protein and keep t...

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