High-level expression of active HIV-1 integrase from a synthetic gene in human cells
High-level expression of active HIV-1 integrase from a synthetic gene in human cells
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DOI：
10.1096/fj.14.10.1389
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发表时间：
2000-07-01
期刊：
影响因子：
4.8
通讯作者：
Debyser, Z
中科院分区：
文献类型：
--
作者：
Cherepanov, P;Pluymers, W;Debyser, Z
设计编码HIV-1整合酶的合成基因是为了规避野生型基因中存在的内在不稳定性和阻遏元件。HIV-1整合酶的高水平表达在各种人类细胞系中获得独立的病毒辅助蛋白。选择稳定表达HIV-1整合酶并具有与亲本细胞系相当的生长动力学的人293 T细胞系。该酶定位于细胞核中，并在有丝分裂过程中与染色体保持稳定的结合。在整合酶基因中携带失活D 64 V突变的慢病毒载体颗粒能够稳定转导293 T细胞，当在生产细胞中用从合成基因表达的整合酶补充时。当用缺陷病毒颗粒感染稳定表达整合酶的细胞系时，也检测到整合酶活性的互补。在人类细胞中表达活性HIV-1整合酶将有助于研究整合过程中宿主和病毒因子之间的相互作用。
A synthetic gene encoding for HIV-1 integase was designed to circumvent the intrinsic instability and the repressor elements present in the wild-type gene. High-level expression of HIV-1 integrase was obtained in various human cell lines independently of viral accessory proteins. A human 293T cell line was selected that stably expresses HIV-1 integrase and has growth kinetics comparable to the parental cell line. The enzyme was localized in the nucleus and remained stably associated with the chromosomes during mitosis. Lentiviral vector particles carrying the inactivating D64V mutation in the integase gene were capable of stably transducing 293T cells when complemented in the producer cells with integrase expressed from the synthetic gene. When the cell line that stably expresses integase was infected with the defective viral particles, complementation of integrase activity was detected as well. Expression of active HIV-1 integase in human cells will facilitate the study of the interplay between host and viral factors during integation.