在 HIV-1 感染过程中核内衣壳的生化检测。

Biochemical detection of capsid in the nucleus during HIV-1 infection.

机构信息

Department of Microbiology and Immunology, Einstein, Bronx, NY 10461, USA.

出版信息

STAR Protoc. 2021 Feb 3;2(1):100323. doi: 10.1016/j.xpro.2021.100323. eCollection 2021 Mar 19.

DOI:10.1016/j.xpro.2021.100323

PMID:33604582

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7873705/

Abstract

To understand the role of the HIV-1 capsid in viral replication, we developed a protocol to biochemically track capsid in the nucleus during infection. To this end, we separated HIV-1-infected cells into nuclear and cytosolic fractions. Fractions were analyzed by western blotting for HIV-1 capsid content as well as for nuclear and cytosolic markers to assess the bona fide origin of the fractions. This protocol can be applied in both cycling and non-cycling human cells. For complete details on the use and execution of this protocol, please refer to Selyutina et al. (2020a).

摘要

为了了解 HIV-1 衣壳在病毒复制中的作用，我们开发了一种生化方法来跟踪感染过程中衣壳在核内的情况。为此，我们将感染 HIV-1 的细胞分离成核和胞质部分。通过 Western blot 分析各部分的 HIV-1 衣壳含量以及核和胞质标记物，以评估各部分的真实来源。该方案可应用于有丝分裂和非有丝分裂的人类细胞。有关此方案的使用和实施的完整详细信息，请参见 Selyutina 等人（2020a）。