SARS-CoV-2 蛋白的荧光构建体综合文库及其在不同细胞类型中的初步特征。
A comprehensive library of fluorescent constructs of SARS-CoV-2 proteins and their initial characterisation in different cell types.
机构信息
Department of Cell Biology and Cancer, Institut Curie, PSL Research University, Sorbonne Université, CNRS, UMR144, Paris, F-75005, France.
Mediterranean Institute for Life Sciences (MedILS), Split, 21000, Croatia.
出版信息
Biol Cell. 2021 Jul;113(7):311-328. doi: 10.1111/boc.202000158. Epub 2021 May 10.
Abstract
BACKGROUND INFORMATION
Comprehensive libraries of plasmids for SARS-CoV-2 proteins with various tags (e.g., Strep, HA, Turbo) are now available. They enable the identification of numerous potential protein-protein interactions between the SARS-CoV-2 virus and host proteins.
RESULTS
We present here a large library of SARS CoV-2 protein constructs fused with green and red fluorescent proteins and their initial characterisation in various human cell lines including lung epithelial cell models (A549, BEAS-2B), as well as in budding yeast. The localisation of a few SARS-CoV-2 proteins matches their proposed interactions with host proteins. These include the localisation of Nsp13 to the centrosome, Orf3a to late endosomes and Orf9b to mitochondria.
CONCLUSIONS AND SIGNIFICANCE
This library should facilitate further cellular investigations, notably by imaging techniques.
摘要
背景信息
现在已经有了包含各种标签（例如 Strep、HA、Turbo）的 SARS-CoV-2 蛋白的综合质粒文库。这些文库能够鉴定出 SARS-CoV-2 病毒与宿主蛋白之间的许多潜在的蛋白-蛋白相互作用。
结果
我们在这里展示了一个大型 SARS-CoV-2 蛋白文库，这些蛋白与绿色和红色荧光蛋白融合，并在各种人类细胞系中进行了初步鉴定，包括肺上皮细胞模型（A549、BEAS-2B）和 budding yeast。一些 SARS-CoV-2 蛋白的定位与其与宿主蛋白的预期相互作用相匹配。这包括 Nsp13 定位于中心体、Orf3a 定位于晚期内体和 Orf9b 定位于线粒体。
结论和意义
这个文库应该有助于进一步的细胞研究，特别是通过成像技术。
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