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[Establishment of C26 cell strain stably expressing folate receptor α]
Zheng Qiu1, Lijun Xing1, Xinxin Liu1
1State Key Laboratory of Natural Medicines, Department of Molecular Biology, College of Life Sciences and Technology, China Pharmaceutical University, Nanjing 210009, China.
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 LipofectamineTM2000. 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 the stability of FRα expression after several generations.
Conclusion:
We have established the cell strain stably expressing FRα, which offer a tool to evaluate the effect of DNA vaccine based on FRα.
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