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Author:
Date:
2016-12-05
[Abstract] Viral pseudotyped particles (pp) are enveloped virus particles, typically derived from retroviruses or rhabdoviruses, that harbor heterologous envelope glycoproteins on their surface and a genome lacking essential genes. These synthetic viral particles are safer surrogates of native viruses and acquire the tropism and host entry pathway characteristics governed by the heterologous envelope glycoprotein used. They have proven to be very useful tools used in research with many applications, such as enabling the study of entry pathways of enveloped viruses and to generate effective gene-delivery vectors. The basis for their generation lies in the capacity of some viruses, such as murine leukemia virus (MLV), to incorporate envelope glycoproteins of other viruses into a pseudotyped virus ...
[摘要] 病毒假型颗粒（pp）是包膜病毒颗粒，通常来自逆转录病毒或弹状病毒，在其表面上携带异源性包膜糖蛋白和缺乏必需基因的基因组。这些合成的病毒颗粒是天然病毒的更安全的替代品并且获得由所使用的异源性包膜糖蛋白控制的嗜性和宿主进入途径特征。它们已被证明是用于具有许多应用的研究中非常有用的工具，例如使得能够研究包膜病毒的进入途径并产生有效的基因递送载体。它们的产生的基础在于一些病毒如鼠白血病病毒（MLV）的能力，以将其他病毒的包膜糖蛋白掺入假型病毒颗粒中。这些可以被工程化以包含报道基因例如荧光素酶，使得能够在易感细胞的假型粒子感染时定量病毒进入事件。在这里，我们详细说明使用中东呼吸综合征冠状病毒（MERS-CoV）尖峰（S）作为待并入的异源包膜糖蛋白的实例，能够产生基于MLV的假型包装颗粒的方案。我们还描述了这些颗粒如何用于感染易感细胞并通过荧光素酶测定进行定量感染性读数。
关键词：假型颗粒，鼠白血病病毒，包膜糖蛋白，冠状病毒，
[背景] ...
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Author:
Date:
2015-08-05
[Abstract] Among the seven serines and one threonine in the carboxyl-terminus of HBV C protein, all but one (serine 183) appear in the context of RxxS/T consensus phosphoacceptor motifs and also overlap with other consensus motifs, such as S/TP, RS, SPRRR, RRRS/T, or RRxS/T, suggesting that various cellular kinases phosphorylate these residues. To determine whether threonine and/or serine (serines 157, 164, 170, 172, 178, and 180, and threonine 162, adw subtype) of HBV C protein are indeed phosphoacceptor residues in cells, Huh7 were transfected with a series of C-protein-expressing mutants, labeled with 32P-orthophosphate for 14 h, and then lysed. The 32Pi-labeled lysates were immunoprecipitated with anti-HBc antibody, and the 32Pi-labeled immunoprecipitated C ...
[摘要] 在HBV C蛋白的羧基末端中的七个丝氨酸和一个苏氨酸中，除了一个（丝氨酸183）外，所有其它丝氨酸出现在RxxS/T共有磷酸受体基序的上下文中，并且还与其他共有基序重叠，例如S/TP，RS ，SPRRR，RRRS/T或RRxS/T，表明各种细胞激酶磷酸化这些残基。 为了确定HBV C蛋白的苏氨酸和/或丝氨酸（丝氨酸157,164,170,172,178和180以及苏氨酸162，adw亚型）是否确实是细胞中的磷酸受体残基，Huh7用一系列C- 蛋白表达突变体，用正磷酸32P标记14小时，然后裂解。 用抗HBc抗体免疫沉淀 32 P标记的裂解物，并通过放射自显影检测32 P标记的免疫沉淀的C蛋白。
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