La Porte Annalena, Kalpana Ganjam V
Department of Genetics, Albert Einstein College ofMedicine, 1300 Morris Park Avenue, Bronx, NY, 10461, USA.
Departments of Genetics, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY, 10461, USA.
Methods Mol Biol. 2016;1354:165-74. doi: 10.1007/978-1-4939-3046-3_11.
Trafficking of newly synthesized Gag protein to the plasma membrane is one of the important steps during HIV-1 assembly. It requires participation of both viral and cellular determinants. Several techniques have been used to measure the amount of Gag that is associated with plasma membrane. Here we describe a microscopy-based method to estimate the distribution of Gag protein within the producer cell. This method can be used in conjunction with other biochemical techniques to quantify the distribution of Gag within a virus-producing cell and its accumulation at the plasma membrane. Since this method is microscopy based, it allows one to quantitate Gag across the cytoplasm, from the nuclear periphery to plasma membrane, at the single-cell level.
新合成的Gag蛋白运输到质膜是HIV-1组装过程中的重要步骤之一。这需要病毒和细胞决定因素的参与。已经使用了几种技术来测量与质膜相关的Gag量。在这里，我们描述了一种基于显微镜的方法来估计Gag蛋白在产生细胞内的分布。该方法可与其他生化技术结合使用，以量化Gag在病毒产生细胞内的分布及其在质膜上的积累。由于该方法基于显微镜，因此它允许在单细胞水平上定量细胞质中从核周边到质膜的Gag。
