Establishment and Characterization of THP-1 Cell Model Stably Expressing HIV-1 Auxiliary Protein Rev
陆小鹏 周慧芳, LU Xiao-peng Zhou Hui-fang
Abstract
陆小鹏 周慧芳, LU Xiao-peng Zhou Hui-fang
Abstract
HIV-1 auxiliary proteins play a critical role in both the infection of HIV-1 target cells and the morbidity of AIDS. Rev, as one of the HIV-1 auxiliary proteins, is essential for the replication of HIV-1. As a membrane-shuttle protein, it can regulate the transportation of HIV-1 structural protein’s mRNA. The rev gene from HIV-1 was transfected into THP-1 cells. The Rev-stably-expressed cell model was generated by electroporation and then sorted and selected with G418. The cell model was characterized at both mRNA and protein levels with an RT-PCR assay, fluorescent observation, and flow cytometry analysis. The rev gene was successfully transfected into THP-1 cells and stably expressed. This is a useful result for future research of the interaction between Rev and cells.
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HIV-1 auxiliary proteins play a critical role in both the infection of HIV-1 target cells and the morbidity of AIDS. Rev, as one of the HIV-1 auxiliary proteins, is essential for the replication of HIV-1. As a membrane-shuttle protein, it can regulate the transportation of HIV-1 structural protein’s mRNA. The rev gene from HIV-1 was transfected into THP-1 cells. The Rev-stably-expressed cell model was generated by electroporation and then sorted and selected with G418. The cell model was characterized at both mRNA and protein levels with an RT-PCR assay, fluorescent observation, and flow cytometry analysis. The rev gene was successfully transfected into THP-1 cells and stably expressed. This is a useful result for future research of the interaction between Rev and cells.
Key concepts: Transfection, Electroporation, Flow cytometry, Molecular biology, Messenger RNA, Human immunodeficiency virus (HIV), Gene, Cell