Open AccessStructural genomics of the SARS coronavirus: cloning, expression, crystallization and preliminary crystallographic study of the Nsp9 protein
Author(s) -
Campanacci Valérie,
Egloff MariePierre,
Longhi Sonia,
Ferron François,
Rancurel Corinne,
Salomoni Aurelia,
Durousseau Cécile,
Tocque Fabienne,
Brémond Nicolas,
Dobbe Jessika C.,
Snijder Eric J.,
Canard Bruno,
Cambillau Christian
Publication year - 2003
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
ISSN - 1399-0047
DOI - 10.1107/s0907444903016779
Subject(s) - cloning (programming) , structural genomics , genome , rna , coronaviridae , crystallization , escherichia coli , biology , virology , coronavirus , virus , gene , chemistry , covid-19 , protein structure , genetics , biochemistry , medicine , organic chemistry , pathology , computer science , infectious disease (medical specialty) , programming language , disease
The aetiologic agent of the recent epidemics of Severe Acute Respiratory Syndrome (SARS) is a positive‐stranded RNA virus (SARS‐CoV) belonging to the Coronaviridae family and its genome differs substantially from those of other known coronaviruses. SARS‐CoV is transmissible mainly by the respiratory route and to date there is no vaccine and no prophylactic or therapeutic treatments against this agent. A SARS‐CoV whole‐genome approach has been developed aimed at determining the crystal structure of all of its proteins or domains. These studies are expected to greatly facilitate drug design. The genomes of coronaviruses are between 27 and 31.5 kbp in length, the largest of the known RNA viruses, and encode 20–30 mature proteins. The functions of many of these polypeptides, including the Nsp9–Nsp10 replicase‐cleavage products, are still unknown. Here, the cloning, Escherichia coli expression, purification and crystallization of the SARS‐CoV Nsp9 protein, the first SARS‐CoV protein to be crystallized, are reported. Nsp9 crystals diffract to 2.8 Å resolution and belong to space group P 6 1/5 22, with unit‐cell parameters a = b = 89.7, c = 136.7 Å. With two molecules in the asymmetric unit, the solvent content is 60% ( V M = 3.1 Å 3 Da −1 ).