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周慧芳, 陆小鹏, 孙正华, 郭 彦, 张华堂. 2008: 稳定表达HIV-1辅助蛋白Rev的THP-1细胞模型的建立和鉴定. 动物学研究, 29(4): 421-426. DOI: 10.3724/SP.J.1141.2008.04421
引用本文: 周慧芳, 陆小鹏, 孙正华, 郭 彦, 张华堂. 2008: 稳定表达HIV-1辅助蛋白Rev的THP-1细胞模型的建立和鉴定. 动物学研究, 29(4): 421-426. DOI: 10.3724/SP.J.1141.2008.04421
ZHOU Hui-fang, LU Xiao-peng, SUN Zheng-hua, GUO Yan, ZHANG Hua-tang. 2008. Establishment and Characterization of THP-1 Cell Model Stably Expressing HIV-1 Auxiliary Protein Rev. Zoological Research, 29(4): 421-426. DOI: 10.3724/SP.J.1141.2008.04421
Citation: ZHOU Hui-fang, LU Xiao-peng, SUN Zheng-hua, GUO Yan, ZHANG Hua-tang. 2008. Establishment and Characterization of THP-1 Cell Model Stably Expressing HIV-1 Auxiliary Protein Rev. Zoological Research, 29(4): 421-426. DOI: 10.3724/SP.J.1141.2008.04421

稳定表达HIV-1辅助蛋白Rev的THP-1细胞模型的建立和鉴定

Establishment and Characterization of THP-1 Cell Model Stably Expressing HIV-1 Auxiliary Protein Rev

  • 摘要: Human immunodeficiency virus-1(HIV-1)辅助蛋白在其感染和艾滋病发病过程中起着非常重要的作用。Regulator of expression of virion proteins(Rev)作为HIV-1辅助蛋白之一,可以调节病毒结构蛋白mRNA出核转运和蛋白表达,对于病毒的复制至关重要。为研究Rev蛋白对靶细胞表型和功能的影响,本实验采用电穿孔的方法,将HIV-1的rev基因导入THP-1细胞,通过流式分选结合G418筛选的方法建立稳定表达Rev蛋白的细胞模型;并通过RT-PCR、荧光观察及流式检测的方法,在mRNA和蛋白两个水平对所建立的细胞模型进行鉴定。结果证实rev基因成功导入了THP-1细胞并稳定表达,为后续rev基因产物与细胞相互作用的研究提供了平台。

     

    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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