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1型人类免疫缺陷病毒整合前复合物中存在的病毒蛋白的测定。

Determination of viral proteins present in the human immunodeficiency virus type 1 preintegration complex.

作者信息

Farnet C M, Haseltine W A

机构信息

Dana-Farber Cancer Institute, Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115.

出版信息

J Virol. 1991 Apr;65(4):1910-5. doi: 10.1128/JVI.65.4.1910-1915.1991.

Abstract

Cytoplasmic extracts prepared from cells infected with metabolically radiolabeled virions of human immunodeficiency virus type 1 contain viral DNA in association with labeled viral proteins. Viral DNA can be purified from these extracts by gel filtration chromatography and sucrose gradient sedimentation as a part of a nucleoprotein complex containing integrase as the only viral protein detectable by immunoprecipitation and gel electrophoretic analysis. The purified complex contains no detectable gag gene products, including p17, p24, p7, or p6, and contains no additional pol gene products, including the p10 protease, p66 and p51 polymerase, or the p15 RNase H. Nearly all of the purified nucleoprotein complexes are capable of integrating into heterologous DNA targets in vitro. These observations demonstrate that integrase is a component of the human immunodeficiency virus type 1 preintegration complex and suggest that integrase may be the only viral protein necessary for the integration of retroviral DNA.

摘要

从感染了用放射性代谢标记的1型人类免疫缺陷病毒病毒体的细胞中制备的细胞质提取物,含有与标记的病毒蛋白相关联的病毒DNA。病毒DNA可通过凝胶过滤色谱法和蔗糖梯度沉降法从这些提取物中纯化出来,作为一种核蛋白复合物的一部分,该复合物包含整合酶,是通过免疫沉淀和凝胶电泳分析唯一可检测到的病毒蛋白。纯化的复合物中未检测到gag基因产物,包括p17、p24、p7或p6,也未检测到其他pol基因产物,包括p10蛋白酶、p66和p51聚合酶或p15核糖核酸酶H。几乎所有纯化的核蛋白复合物都能够在体外整合到异源DNA靶标中。这些观察结果表明,整合酶是1型人类免疫缺陷病毒整合前复合物的一个组成部分,并提示整合酶可能是逆转录病毒DNA整合所必需的唯一病毒蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d8ec/240011/3397d7b6f19d/jvirol00047-0256-a.jpg

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