Genetic screen for monitoring severe acute respiratory syndrome coronavirus 3C-like protease.
Identifieur interne : 000C33 ( Ncbi/Curation ); précédent : 000C32; suivant : 000C34Genetic screen for monitoring severe acute respiratory syndrome coronavirus 3C-like protease.
Auteurs : Mariona Parera [Espagne] ; Bonaventura Clotet ; Miguel Angel MartinezSource :
- Journal of virology [ 0022-538X ] ; 2004.
Descripteurs français
- KwdFr :
- Analyse de séquence d'ADN, Bactériophage lambda (génétique), Cysteine endopeptidases (), Cysteine endopeptidases (génétique), Cysteine endopeptidases (métabolisme), Cysteine endopeptidases (synthèse chimique), Données de séquences moléculaires, Humains, Syndrome respiratoire aigu sévère (virologie), Séquence d'acides aminés, Techniques génétiques, Virus du SRAS (enzymologie), Virus du SRAS (génétique).
- MESH :
- enzymologie : Virus du SRAS.
- génétique : Bactériophage lambda, Cysteine endopeptidases, Virus du SRAS.
- métabolisme : Cysteine endopeptidases.
- synthèse chimique : Cysteine endopeptidases.
- virologie : Syndrome respiratoire aigu sévère.
- Analyse de séquence d'ADN, Cysteine endopeptidases, Données de séquences moléculaires, Humains, Séquence d'acides aminés, Techniques génétiques.
English descriptors
- KwdEn :
- Amino Acid Sequence, Bacteriophage lambda (genetics), Cysteine Endopeptidases (chemical synthesis), Cysteine Endopeptidases (chemistry), Cysteine Endopeptidases (genetics), Cysteine Endopeptidases (metabolism), Genetic Techniques, Humans, Molecular Sequence Data, SARS Virus (enzymology), SARS Virus (genetics), Sequence Analysis, DNA, Severe Acute Respiratory Syndrome (virology).
- MESH :
- chemical , chemical synthesis : Cysteine Endopeptidases.
- chemical , chemistry : Cysteine Endopeptidases.
- enzymology : SARS Virus.
- genetics : Bacteriophage lambda, Cysteine Endopeptidases, SARS Virus.
- chemical , metabolism : Cysteine Endopeptidases.
- virology : Severe Acute Respiratory Syndrome.
- Amino Acid Sequence, Genetic Techniques, Humans, Molecular Sequence Data, Sequence Analysis, DNA.
Abstract
A novel coronavirus (SCoV) is the etiological agent of severe acute respiratory syndrome. Site-specific proteolysis plays a critical role in regulating a number of cellular and viral processes. Since the main protease of SCoV, also termed 3C-like protease, is an attractive target for drug therapy, we have developed a safe, simple, and rapid genetic screen assay to monitor the activity of the SCoV 3C-like protease. This genetic system is based on the bacteriophage lambda regulatory circuit, in which the viral repressor cI is specifically cleaved to initiate the lysogenic-to-lytic switch. A specific target for the SCoV 3C-like protease, P1/P2 (SAVLQ/SGFRK), was inserted into the lambda phage cI repressor. The target specificity of the SCoV P1/P2 repressor was evaluated by coexpression of this repressor with a chemically synthesized SCoV 3C-like protease gene construct. Upon infection of Escherichia coli cells containing the two plasmids encoding the cI. SCoV P1/P2-cro and the beta-galactosidase-SCoV 3C-like protease constructs, lambda phage replicated up to 2,000-fold more efficiently than in cells that did not express the SCoV 3C-like protease. This simple and highly specific assay can be used to monitor the activity of the SCoV 3C-like protease, and it has the potential to be used for screening specific inhibitors.
DOI: 10.1128/JVI.78.24.14057-14061.2004
PubMed: 15564515
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chimique)</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Séquence d'acides aminés</term><term>Techniques génétiques</term><term>Virus du SRAS (enzymologie)</term><term>Virus du SRAS (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemical synthesis" xml:lang="en"><term>Cysteine Endopeptidases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Cysteine Endopeptidases</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Bacteriophage lambda</term><term>Cysteine Endopeptidases</term><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Bactériophage lambda</term><term>Cysteine endopeptidases</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Cysteine Endopeptidases</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Cysteine endopeptidases</term></keywords><keywords scheme="MESH" qualifier="synthèse chimique" xml:lang="fr"><term>Cysteine endopeptidases</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Genetic Techniques</term><term>Humans</term><term>Molecular Sequence Data</term><term>Sequence Analysis, DNA</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Analyse de séquence d'ADN</term><term>Cysteine endopeptidases</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Séquence d'acides aminés</term><term>Techniques génétiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A novel coronavirus (SCoV) is the etiological agent of severe acute respiratory syndrome. Site-specific proteolysis plays a critical role in regulating a number of cellular and viral processes. Since the main protease of SCoV, also termed 3C-like protease, is an attractive target for drug therapy, we have developed a safe, simple, and rapid genetic screen assay to monitor the activity of the SCoV 3C-like protease. This genetic system is based on the bacteriophage lambda regulatory circuit, in which the viral repressor cI is specifically cleaved to initiate the lysogenic-to-lytic switch. A specific target for the SCoV 3C-like protease, P1/P2 (SAVLQ/SGFRK), was inserted into the lambda phage cI repressor. The target specificity of the SCoV P1/P2 repressor was evaluated by coexpression of this repressor with a chemically synthesized SCoV 3C-like protease gene construct. Upon infection of Escherichia coli cells containing the two plasmids encoding the cI. SCoV P1/P2-cro and the beta-galactosidase-SCoV 3C-like protease constructs, lambda phage replicated up to 2,000-fold more efficiently than in cells that did not express the SCoV 3C-like protease. This simple and highly specific assay can be used to monitor the activity of the SCoV 3C-like protease, and it has the potential to be used for screening specific inhibitors.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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