Expression, purification and characterization of recombinant severe acute respiratory syndrome coronavirus non-structural protein 1.
Identifieur interne : 003704 ( Main/Exploration ); précédent : 003703; suivant : 003705Expression, purification and characterization of recombinant severe acute respiratory syndrome coronavirus non-structural protein 1.
Auteurs : Kimberly Brucz [États-Unis] ; Zachary J. Miknis ; L Wayne Schultz ; Timothy C. UmlandSource :
- Protein expression and purification [ 1046-5928 ] ; 2007.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : Recombinant Proteins.
- chemical , metabolism : Peptide Hydrolases, Recombinant Proteins.
- chemistry : SARS Virus.
- genetics : SARS Virus.
- Chromatography, Gel, Circular Dichroism, Gene Expression, Molecular Weight, Viral Nonstructural Proteins.
Abstract
The coronavirus (CoV) responsible for severe acute respiratory syndrome (SARS), SARS-CoV, encodes two large polyproteins (pp1a and pp1ab) that are processed by two viral proteases to yield mature non-structural proteins (nsps). Many of these nsps have essential roles in viral replication, but several have no assigned function and possess amino acid sequences that are unique to the CoV family. One such protein is SARS-CoV nsp1, which is processed from the N-terminus of both pp1a and pp1ab. The mature SARS-CoV protein is present in cells several hours post-infection and co-localizes to the viral replication complex, but its function in the viral life cycle remains unknown. Furthermore, nsp1 sequences are highly divergent across the CoV family, and it has been suggested that this is due to nsp1 possessing a function specific to viral interactions with its host cell or acting as a host specific virulence factor. In order to initiate structural and biophysical studies of SARS-CoV nsp1, a recombinant expression system and a purification protocol have been developed, yielding milligram quantities of highly purified SARS-CoV nsp1. The purified protein was characterized using circular dichroism, size exclusion chromatography, and multi-angle light scattering.
DOI: 10.1016/j.pep.2006.11.005
PubMed: 17187987
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: 001F41
- to stream PubMed, to step Curation: 001F41
- to stream PubMed, to step Checkpoint: 001E09
- to stream Ncbi, to step Merge: 001819
- to stream Ncbi, to step Curation: 001819
- to stream Ncbi, to step Checkpoint: 001819
- to stream Main, to step Merge: 003845
- to stream Main, to step Curation: 003704
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Expression, purification and characterization of recombinant severe acute respiratory syndrome coronavirus non-structural protein 1.</title><author><name sortKey="Brucz, Kimberly" sort="Brucz, Kimberly" uniqKey="Brucz K" first="Kimberly" last="Brucz">Kimberly Brucz</name><affiliation wicri:level="2"><nlm:affiliation>Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, NY 14203, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, NY 14203</wicri:regionArea><placeName><region type="state">État de New York</region></placeName></affiliation></author><author><name sortKey="Miknis, Zachary J" sort="Miknis, Zachary J" uniqKey="Miknis Z" first="Zachary J" last="Miknis">Zachary J. Miknis</name></author><author><name sortKey="Schultz, L Wayne" sort="Schultz, L Wayne" uniqKey="Schultz L" first="L Wayne" last="Schultz">L Wayne Schultz</name></author><author><name sortKey="Umland, Timothy C" sort="Umland, Timothy C" uniqKey="Umland T" first="Timothy C" last="Umland">Timothy C. Umland</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2007">2007</date><idno type="RBID">pubmed:17187987</idno><idno type="pmid">17187987</idno><idno type="doi">10.1016/j.pep.2006.11.005</idno><idno type="wicri:Area/PubMed/Corpus">001F41</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001F41</idno><idno type="wicri:Area/PubMed/Curation">001F41</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001F41</idno><idno type="wicri:Area/PubMed/Checkpoint">001E09</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001E09</idno><idno type="wicri:Area/Ncbi/Merge">001819</idno><idno type="wicri:Area/Ncbi/Curation">001819</idno><idno type="wicri:Area/Ncbi/Checkpoint">001819</idno><idno type="wicri:doubleKey">1046-5928:2007:Brucz K:expression:purification:and</idno><idno type="wicri:Area/Main/Merge">003845</idno><idno type="wicri:Area/Main/Curation">003704</idno><idno type="wicri:Area/Main/Exploration">003704</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Expression, purification and characterization of recombinant severe acute respiratory syndrome coronavirus non-structural protein 1.</title><author><name sortKey="Brucz, Kimberly" sort="Brucz, Kimberly" uniqKey="Brucz K" first="Kimberly" last="Brucz">Kimberly Brucz</name><affiliation wicri:level="2"><nlm:affiliation>Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, NY 14203, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Hauptman-Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, NY 14203</wicri:regionArea><placeName><region type="state">État de New York</region></placeName></affiliation></author><author><name sortKey="Miknis, Zachary J" sort="Miknis, Zachary J" uniqKey="Miknis Z" first="Zachary J" last="Miknis">Zachary J. Miknis</name></author><author><name sortKey="Schultz, L Wayne" sort="Schultz, L Wayne" uniqKey="Schultz L" first="L Wayne" last="Schultz">L Wayne Schultz</name></author><author><name sortKey="Umland, Timothy C" sort="Umland, Timothy C" uniqKey="Umland T" first="Timothy C" last="Umland">Timothy C. Umland</name></author></analytic><series><title level="j">Protein expression and purification</title><idno type="ISSN">1046-5928</idno><imprint><date when="2007" type="published">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Chromatography, Gel</term><term>Circular Dichroism</term><term>Gene Expression</term><term>Molecular Weight</term><term>Peptide Hydrolases (metabolism)</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (metabolism)</term><term>SARS Virus (chemistry)</term><term>SARS Virus (genetics)</term><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Chromatographie sur gel</term><term>Dichroïsme circulaire</term><term>Expression des gènes</term><term>Masse moléculaire</term><term>Peptide hydrolases (métabolisme)</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (métabolisme)</term><term>Protéines virales non structurales</term><term>Virus du SRAS ()</term><term>Virus du SRAS (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Peptide Hydrolases</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines recombinantes</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Peptide hydrolases</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Chromatography, Gel</term><term>Circular Dichroism</term><term>Gene Expression</term><term>Molecular Weight</term><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Chromatographie sur gel</term><term>Dichroïsme circulaire</term><term>Expression des gènes</term><term>Masse moléculaire</term><term>Protéines virales non structurales</term><term>Virus du SRAS</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The coronavirus (CoV) responsible for severe acute respiratory syndrome (SARS), SARS-CoV, encodes two large polyproteins (pp1a and pp1ab) that are processed by two viral proteases to yield mature non-structural proteins (nsps). Many of these nsps have essential roles in viral replication, but several have no assigned function and possess amino acid sequences that are unique to the CoV family. One such protein is SARS-CoV nsp1, which is processed from the N-terminus of both pp1a and pp1ab. The mature SARS-CoV protein is present in cells several hours post-infection and co-localizes to the viral replication complex, but its function in the viral life cycle remains unknown. Furthermore, nsp1 sequences are highly divergent across the CoV family, and it has been suggested that this is due to nsp1 possessing a function specific to viral interactions with its host cell or acting as a host specific virulence factor. In order to initiate structural and biophysical studies of SARS-CoV nsp1, a recombinant expression system and a purification protocol have been developed, yielding milligram quantities of highly purified SARS-CoV nsp1. The purified protein was characterized using circular dichroism, size exclusion chromatography, and multi-angle light scattering.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>État de New York</li></region></list><tree><noCountry><name sortKey="Miknis, Zachary J" sort="Miknis, Zachary J" uniqKey="Miknis Z" first="Zachary J" last="Miknis">Zachary J. Miknis</name><name sortKey="Schultz, L Wayne" sort="Schultz, L Wayne" uniqKey="Schultz L" first="L Wayne" last="Schultz">L Wayne Schultz</name><name sortKey="Umland, Timothy C" sort="Umland, Timothy C" uniqKey="Umland T" first="Timothy C" last="Umland">Timothy C. Umland</name></noCountry><country name="États-Unis"><region name="État de New York"><name sortKey="Brucz, Kimberly" sort="Brucz, Kimberly" uniqKey="Brucz K" first="Kimberly" last="Brucz">Kimberly Brucz</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/SrasV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 003704 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 003704 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= SrasV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= pubmed:17187987
|texte= Expression, purification and characterization of recombinant severe acute respiratory syndrome coronavirus non-structural protein 1.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:17187987" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a SrasV1
| This area was generated with Dilib version V0.6.33. |  |