| [14C] Linoleic Acid Uptake and Fractionation Assay in Vibrio cholerae
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Author:
Date:
2015-12-20
[Abstract] The gram-negative curved bacillus Vibrio cholerae (V. cholerae) causes the severe diarrheal illness cholera. The work presented here is to assess whether unsaturated fatty acids (UFAs), such as linoleic acid, have the potential to directly affect proteins involved in DNA binding because they are able to enter the cell. In this protocol, we show how to measure linoleic acid entering V. cholerae when added exogenously and determine whether it is able to enter the cytoplasm. This protocol will quantify how much linoleic acid is able to enter the cell and then identify the amount of linoleic acid that stays in the membrane or ultimately enters the cytoplasm.
[摘要] 革兰氏阴性弯曲杆菌霍乱弧菌（<霍乱弧菌）导致严重的腹泻病霍乱。 这里介绍的工作是评估是否不饱和脂肪酸（ufas），如亚油酸，有可能直接影响参与dna结合的蛋白质，因为他们能够进入细胞。 在这个协议，我们显示如何测量亚麻酸进入。 霍乱菌，并且确定它是否能够进入细胞质。 该方案将量化亚油酸能够进入细胞多少，然后鉴定保留在膜中或最终进入细胞质的亚油酸的量。
这里介绍的工作是评估是否不饱和脂肪酸（ufas），如亚油酸，有可能直接影响参与dna结合的蛋白质，因为他们能够进入细胞。="" 在这个协议，我们显示如何测量亚麻酸进入。="" 霍乱菌，并且确定它是否能够进入细胞质。="" 该方案将量化亚油酸能够进入细胞多少，然后鉴定保留在膜中或最终进入细胞质的亚油酸的量。="">
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| HIV-1 Virus-like Particle Budding Assay
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Author:
Date:
2013-07-20
[Abstract] Viral replication culminates with the egress of the mature virion from the host cell. This step of the viral life cycle has recently garnered increased attention with the discovery of the cellular restriction factor, Tetherin, which tethers budded virions to the surface of infected cells and inhibits viral spread. The importance of this block in viral infections has been suggested by the discovery of viral antagonists, such as HIV-1 Vpu, which counteract Tetherin. This protocol describes a system to study HIV-1 budding under BSL-2 safety conditions. It takes advantage of the ability of many viral matrix/capsid proteins to generate non-infectious virus-like particles (VLPs) with the expression of a single viral protein (i.e. HIV-1 p24 Gag). This protocol was recently used to ...
[摘要] 病毒复制随着来自宿主细胞的成熟病毒体的流出而达到高潮。病毒生命周期的这个步骤最近通过发现细胞限制因子Tetherin而发现增加的注意力，Tetherin将暴露的病毒粒子束缚于感染细胞的表面并抑制病毒扩散。这种阻断在病毒感染中的重要性已经通过发现病毒拮抗剂如HIV-1 Vpu（其抵抗了细胞因子）而提出。该协议描述了在BSL-2安全条件下研究HIV-1芽生的系统。它利用了许多病毒基质/衣壳蛋白产生具有单个病毒蛋白（即HIV-1p24Gag）表达的非感染性病毒样颗粒（VLPs）的能力。这个协议最近用于表征在HIV-1 Vpu存在下，Tetherin亚型对VLP释放的影响（Cocka和Bates，2012）。同时表达的Tetherin和其他病毒拮抗剂可用于研究Tetherin介导的病毒芽出的限制。
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