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  • A comprehensive library of fluorescent constructs of SARS-CoV-2 proteins and their initial characterisation in different cell types.

A comprehensive library of fluorescent constructs of SARS-CoV-2 proteins and their initial characterisation in different cell types.

Biology of the cell (2021-03-06)
Stéphanie Miserey-Lenkei, Katarina Trajkovic, Juan Martín D'Ambrosio, Amanda J Patel, Alenka Čopič, Pallavi Mathur, Kristine Schauer, Bruno Goud, Véronique Albanèse, Romain Gautier, Melody Subra, David Kovacs, Hélène Barelli, Bruno Antonny
ABSTRACT

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. 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. This library should facilitate further cellular investigations, notably by imaging techniques.

MATERIALS
Product Number
Brand
Product Description

Roche
Anti-GFP, from mouse IgG1κ (clones 7.1 and 13.1)
Sigma-Aldrich
Monoclonal Anti-HSP90B1 antibody produced in mouse, Prestige Antibodies® Powered by Atlas Antibodies, clone CL2647, purified immunoglobulin, buffered aqueous glycerol solution