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首页> 外文期刊>Virology >Nuclear localization of human immunodeficiency virus type 1 integrase expressed as a fusion protein with green fluorescent protein.
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Nuclear localization of human immunodeficiency virus type 1 integrase expressed as a fusion protein with green fluorescent protein.

机译:人类免疫缺陷病毒1型整合酶的核定位表示为与绿色荧光蛋白的融合蛋白。

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摘要

Lentiviruses in general and the human immunodeficiency virus type 1 (HIV-1) in particular have the ability to integrate their genome stably into the chromosome of nondividing cells. Integration of HIV cDNA is mediated by the viral integrase (IN). Apart from its catalytic activity, this enzyme seems to play an important role in the transport of the HIV preintegration complex into the nucleus of nondividing cells. We studied the karyophilic properties of IN by constructing an N-terminal fusion protein of HIV-1 integrase and green fluorescent protein (GFP-IN). Transient expression of GFP-IN in various mammalian cell lines was demonstrated by fluorescence microscopy, flow cytometry, and Western blotting. Although wild-type GFP was localized throughout the cell, GFP-IN was localized predominantly in the nucleus. Nuclear localization of GFP-IN was also obtained after transient transfection of the cells arrested in the G1/S phase of the cell cycle. These results provide compelling evidence for the karyophilic properties of the HIV-1 integrase. Copyright 1999 Academic Press.
机译:一般而言,慢病毒,特别是1型人类免疫缺陷病毒(HIV-1)具有将其基因组稳定整合到非分裂细胞染色体中的能力。 HIV cDNA的整合是由病毒整合酶(IN)介导的。除了其催化活性外,该酶似乎在将HIV预整合复合物转运到非分裂细胞核中起重要作用。我们通过构建HIV-1整合酶和绿色荧光蛋白(GFP-IN)的N端融合蛋白来研究IN的亲核特性。 GFP-IN在各种哺乳动物细胞系中的瞬时表达已通过荧光显微镜,流式细胞术和Western印迹进行了证明。尽管野生型GFP位于整个细胞中,但GFP-IN主要位于细胞核中。瞬时转染停滞在细胞周期G1 / S期的细胞后,也获得了GFP-IN的核定位。这些结果为HIV-1整合酶的亲核特性提供了令人信服的证据。版权所有1999,学术出版社。

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