Expression of SARS-coronavirus nucleocapsid protein in Escherichia coli and Lactococcus lactis for serodiagnosis and mucosal vaccination.
Identifieur interne : 004834 ( Main/Exploration ); précédent : 004833; suivant : 004835Expression of SARS-coronavirus nucleocapsid protein in Escherichia coli and Lactococcus lactis for serodiagnosis and mucosal vaccination.
Auteurs : Huadong Pei [République populaire de Chine] ; Jingfang Liu ; Yun Cheng ; Chaomin Sun ; Chen Wang ; Yueping Lu ; Jie Ding ; Jian Zhou ; Hua XiangSource :
- Applied microbiology and biotechnology [ 0175-7598 ] ; 2005.
Descripteurs français
- KwdFr :
- Administration par voie nasale, Administration par voie orale, Animaux, Anticorps antiviraux (sang), Escherichia coli (métabolisme), Femelle, Glutathione transferase (génétique), Glutathione transferase (métabolisme), Humains, Immunoglobuline G (sang), Lactococcus lactis (métabolisme), Protéines nucléocapside (génétique), Protéines nucléocapside (immunologie), Protéines nucléocapside (métabolisme), Protéines recombinantes (biosynthèse), Protéines recombinantes (immunologie), Protéines recombinantes (isolement et purification), Souris, Souris de lignée BALB C, Spécificité des anticorps, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (immunologie), Tests sérologiques, Vaccination, Vaccins antiviraux (administration et posologie), Vaccins antiviraux (immunologie), Vaccins antiviraux (isolement et purification), Vaccins synthétiques (administration et posologie), Vaccins synthétiques (immunologie), Virus du SRAS (immunologie).
- MESH :
- administration et posologie : Vaccins antiviraux, Vaccins synthétiques.
- biosynthèse : Protéines recombinantes.
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Glutathione transferase, Protéines nucléocapside.
- immunologie : Protéines nucléocapside, Protéines recombinantes, Syndrome respiratoire aigu sévère, Vaccins antiviraux, Vaccins synthétiques, Virus du SRAS.
- isolement et purification : Protéines recombinantes, Vaccins antiviraux.
- métabolisme : Escherichia coli, Glutathione transferase, Lactococcus lactis, Protéines nucléocapside.
- sang : Anticorps antiviraux, Immunoglobuline G.
- Administration par voie nasale, Administration par voie orale, Animaux, Femelle, Humains, Souris, Souris de lignée BALB C, Spécificité des anticorps, Tests sérologiques, Vaccination.
English descriptors
- KwdEn :
- Administration, Intranasal, Administration, Oral, Animals, Antibodies, Viral (blood), Antibody Specificity, Escherichia coli (metabolism), Female, Glutathione Transferase (genetics), Glutathione Transferase (metabolism), Humans, Immunoglobulin G (blood), Lactococcus lactis (metabolism), Mice, Mice, Inbred BALB C, Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (immunology), Nucleocapsid Proteins (metabolism), Recombinant Proteins (biosynthesis), Recombinant Proteins (immunology), Recombinant Proteins (isolation & purification), SARS Virus (immunology), Serologic Tests, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (immunology), Vaccination, Vaccines, Synthetic (administration & dosage), Vaccines, Synthetic (immunology), Viral Vaccines (administration & dosage), Viral Vaccines (immunology), Viral Vaccines (isolation & purification).
- MESH :
- chemical , administration & dosage : Vaccines, Synthetic, Viral Vaccines.
- chemical , biosynthesis : Recombinant Proteins.
- chemical , blood : Antibodies, Viral, Immunoglobulin G.
- chemical , genetics : Glutathione Transferase, Nucleocapsid Proteins.
- chemical , immunology : Nucleocapsid Proteins, Recombinant Proteins, Vaccines, Synthetic, Viral Vaccines.
- chemical , isolation & purification : Recombinant Proteins, Viral Vaccines.
- diagnosis : Severe Acute Respiratory Syndrome.
- immunology : SARS Virus, Severe Acute Respiratory Syndrome.
- metabolism : Escherichia coli, Glutathione Transferase, Lactococcus lactis, Nucleocapsid Proteins.
- Administration, Intranasal, Administration, Oral, Animals, Antibody Specificity, Female, Humans, Mice, Mice, Inbred BALB C, Serologic Tests, Vaccination.
Abstract
The nucleocapsid (N) protein of the severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) is an important antigen for the early diagnosis of SARS and the development of vaccines. It was expressed in Escherichia coli as a fusion with human glutathione S-transferase (hGST) and was confirmed by Western blotting analysis. This recombinant N protein (hGST-N) was purified and used to measure the SARS-CoV N-specific antibody in the sera of eight SARS patients by enzyme-linked immunosorbent assay. Specific antibody response to this purified recombinant N protein was 100% positive in the SARS patients' sera, while none of the control sera from 30 healthy people gave a positive reaction in the same assay. The SARS-CoV N protein was also expressed in Lactococcus lactis in the cytoplasm or secreted into the medium. The N-producing strain MG1363/pSECN and the purified hGST-N protein were respectively administered to mice, either orally or intranasally. Results indicated that orally delivered MG1363/pSECN induced significant N-specific IgG in the sera. In conclusion, our work provides a novel strategy to produce the SARS-CoV N protein for serodiagnosis and for L. lactis-based mucosal vaccines.
DOI: 10.1007/s00253-004-1869-y
PubMed: 15660214
Affiliations:
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sort="Ding, Jie" uniqKey="Ding J" first="Jie" last="Ding">Jie Ding</name></author><author><name sortKey="Zhou, Jian" sort="Zhou, Jian" uniqKey="Zhou J" first="Jian" last="Zhou">Jian Zhou</name></author><author><name sortKey="Xiang, Hua" sort="Xiang, Hua" uniqKey="Xiang H" first="Hua" last="Xiang">Hua Xiang</name></author></analytic><series><title level="j">Applied microbiology and biotechnology</title><idno type="ISSN">0175-7598</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Administration, Intranasal</term><term>Administration, Oral</term><term>Animals</term><term>Antibodies, Viral (blood)</term><term>Antibody Specificity</term><term>Escherichia coli (metabolism)</term><term>Female</term><term>Glutathione Transferase (genetics)</term><term>Glutathione Transferase (metabolism)</term><term>Humans</term><term>Immunoglobulin G (blood)</term><term>Lactococcus lactis (metabolism)</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Nucleocapsid Proteins (genetics)</term><term>Nucleocapsid Proteins (immunology)</term><term>Nucleocapsid Proteins (metabolism)</term><term>Recombinant Proteins (biosynthesis)</term><term>Recombinant Proteins (immunology)</term><term>Recombinant Proteins (isolation & purification)</term><term>SARS Virus (immunology)</term><term>Serologic Tests</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe Acute Respiratory Syndrome (immunology)</term><term>Vaccination</term><term>Vaccines, Synthetic (administration & dosage)</term><term>Vaccines, Synthetic (immunology)</term><term>Viral Vaccines (administration & dosage)</term><term>Viral Vaccines (immunology)</term><term>Viral Vaccines (isolation & purification)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Administration par voie nasale</term><term>Administration par voie 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(immunologie)</term><term>Vaccins antiviraux (isolement et purification)</term><term>Vaccins synthétiques (administration et posologie)</term><term>Vaccins synthétiques (immunologie)</term><term>Virus du SRAS (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="administration & dosage" xml:lang="en"><term>Vaccines, Synthetic</term><term>Viral Vaccines</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Antibodies, Viral</term><term>Immunoglobulin G</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Glutathione Transferase</term><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Nucleocapsid Proteins</term><term>Recombinant Proteins</term><term>Vaccines, 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recombinantes</term><term>Syndrome respiratoire aigu sévère</term><term>Vaccins antiviraux</term><term>Vaccins synthétiques</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Protéines recombinantes</term><term>Vaccins antiviraux</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Escherichia coli</term><term>Glutathione Transferase</term><term>Lactococcus lactis</term><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Escherichia coli</term><term>Glutathione transferase</term><term>Lactococcus lactis</term><term>Protéines nucléocapside</term></keywords><keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>Anticorps antiviraux</term><term>Immunoglobuline G</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Administration, Intranasal</term><term>Administration, Oral</term><term>Animals</term><term>Antibody Specificity</term><term>Female</term><term>Humans</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Serologic Tests</term><term>Vaccination</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Administration par voie nasale</term><term>Administration par voie orale</term><term>Animaux</term><term>Femelle</term><term>Humains</term><term>Souris</term><term>Souris de lignée BALB C</term><term>Spécificité des anticorps</term><term>Tests sérologiques</term><term>Vaccination</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The nucleocapsid (N) protein of the severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) is an important antigen for the early diagnosis of SARS and the development of vaccines. It was expressed in Escherichia coli as a fusion with human glutathione S-transferase (hGST) and was confirmed by Western blotting analysis. This recombinant N protein (hGST-N) was purified and used to measure the SARS-CoV N-specific antibody in the sera of eight SARS patients by enzyme-linked immunosorbent assay. Specific antibody response to this purified recombinant N protein was 100% positive in the SARS patients' sera, while none of the control sera from 30 healthy people gave a positive reaction in the same assay. The SARS-CoV N protein was also expressed in Lactococcus lactis in the cytoplasm or secreted into the medium. The N-producing strain MG1363/pSECN and the purified hGST-N protein were respectively administered to mice, either orally or intranasally. Results indicated that orally delivered MG1363/pSECN induced significant N-specific IgG in the sera. In conclusion, our work provides a novel strategy to produce the SARS-CoV N protein for serodiagnosis and for L. lactis-based mucosal vaccines.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Cheng, Yun" sort="Cheng, Yun" uniqKey="Cheng Y" first="Yun" last="Cheng">Yun Cheng</name><name sortKey="Ding, Jie" sort="Ding, Jie" uniqKey="Ding J" first="Jie" last="Ding">Jie Ding</name><name sortKey="Liu, Jingfang" sort="Liu, Jingfang" uniqKey="Liu J" first="Jingfang" last="Liu">Jingfang Liu</name><name sortKey="Lu, Yueping" sort="Lu, Yueping" uniqKey="Lu Y" first="Yueping" last="Lu">Yueping Lu</name><name sortKey="Sun, Chaomin" sort="Sun, Chaomin" uniqKey="Sun C" first="Chaomin" last="Sun">Chaomin Sun</name><name sortKey="Wang, Chen" sort="Wang, Chen" uniqKey="Wang C" first="Chen" last="Wang">Chen Wang</name><name sortKey="Xiang, Hua" sort="Xiang, Hua" uniqKey="Xiang H" first="Hua" last="Xiang">Hua Xiang</name><name sortKey="Zhou, Jian" sort="Zhou, Jian" uniqKey="Zhou J" first="Jian" last="Zhou">Jian Zhou</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Pei, Huadong" sort="Pei, Huadong" uniqKey="Pei H" first="Huadong" last="Pei">Huadong Pei</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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