Expression of SARS-coronavirus nucleocapsid protein in Escherichia coli and Lactococcus lactis for serodiagnosis and mucosal vaccination
Identifieur interne : 005169 ( Main/Merge ); précédent : 005168; suivant : 005170Expression 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 [République populaire de Chine] ; YUN CHENG [République populaire de Chine] ; CHAOMIN SUN [République populaire de Chine] ; CHEN WANG [République populaire de Chine] ; YUEPING LU [République populaire de Chine] ; JIE DING [République populaire de Chine] ; JIAN ZHOU [République populaire de Chine] ; HUA XIANG [République populaire de Chine]Source :
- Applied microbiology and biotechnology [ 0175-7598 ] ; 2005.
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- Pascal (Inist)
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- topic : Vaccination.
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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.
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en" level="a">Expression of SARS-coronavirus nucleocapsid protein in Escherichia coli and Lactococcus lactis for serodiagnosis and mucosal vaccination</title><author><name sortKey="Huadong Pei" sort="Huadong Pei" uniqKey="Huadong Pei" last="Huadong Pei">HUADONG PEI</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Graduate school of the Chinese Academy of Sciences</s1><s2>Beijing, 100039</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Graduate school of the Chinese Academy of Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Jingfang Liu" sort="Jingfang Liu" uniqKey="Jingfang Liu" last="Jingfang Liu">JINGFANG LIU</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Graduate school of the Chinese Academy of Sciences</s1><s2>Beijing, 100039</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Graduate school of the Chinese Academy of Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Yun Cheng" sort="Yun Cheng" uniqKey="Yun Cheng" last="Yun Cheng">YUN CHENG</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation></author><author><name sortKey="Chaomin Sun" sort="Chaomin Sun" uniqKey="Chaomin Sun" last="Chaomin Sun">CHAOMIN SUN</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Graduate school of the Chinese Academy of Sciences</s1><s2>Beijing, 100039</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Graduate school of the Chinese Academy of Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Chen Wang" sort="Chen Wang" uniqKey="Chen Wang" last="Chen Wang">CHEN WANG</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</s1><s2>Beijing, 100020</s2><s3>CHN</s3><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Yueping Lu" sort="Yueping Lu" uniqKey="Yueping Lu" last="Yueping Lu">YUEPING LU</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</s1><s2>Beijing, 100020</s2><s3>CHN</s3><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Jie Ding" sort="Jie Ding" uniqKey="Jie Ding" last="Jie Ding">JIE DING</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</s1><s2>Beijing, 100020</s2><s3>CHN</s3><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Jian Zhou" sort="Jian Zhou" uniqKey="Jian Zhou" last="Jian Zhou">JIAN ZHOU</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation></author><author><name sortKey="Hua Xiang" sort="Hua Xiang" uniqKey="Hua Xiang" last="Hua Xiang">HUA XIANG</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">06-0009448</idno><date when="2005">2005</date><idno type="stanalyst">PASCAL 06-0009448 INIST</idno><idno type="RBID">Pascal:06-0009448</idno><idno type="wicri:Area/PascalFrancis/Corpus">000569</idno><idno type="wicri:Area/PascalFrancis/Curation">000421</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000649</idno><idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000649</idno><idno type="wicri:doubleKey">0175-7598:2005:Huadong Pei:expression:of:sars</idno><idno type="wicri:Area/Main/Merge">005169</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Expression of SARS-coronavirus nucleocapsid protein in Escherichia coli and Lactococcus lactis for serodiagnosis and mucosal vaccination</title><author><name sortKey="Huadong Pei" sort="Huadong Pei" uniqKey="Huadong Pei" last="Huadong Pei">HUADONG PEI</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Graduate school of the Chinese Academy of Sciences</s1><s2>Beijing, 100039</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Graduate school of the Chinese Academy of Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Jingfang Liu" sort="Jingfang Liu" uniqKey="Jingfang Liu" last="Jingfang Liu">JINGFANG LIU</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Graduate school of the Chinese Academy of Sciences</s1><s2>Beijing, 100039</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Graduate school of the Chinese Academy of Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Yun Cheng" sort="Yun Cheng" uniqKey="Yun Cheng" last="Yun Cheng">YUN CHENG</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation></author><author><name sortKey="Chaomin Sun" sort="Chaomin Sun" uniqKey="Chaomin Sun" last="Chaomin Sun">CHAOMIN SUN</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Graduate school of the Chinese Academy of Sciences</s1><s2>Beijing, 100039</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Graduate school of the Chinese Academy of Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Chen Wang" sort="Chen Wang" uniqKey="Chen Wang" last="Chen Wang">CHEN WANG</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</s1><s2>Beijing, 100020</s2><s3>CHN</s3><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Yueping Lu" sort="Yueping Lu" uniqKey="Yueping Lu" last="Yueping Lu">YUEPING LU</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</s1><s2>Beijing, 100020</s2><s3>CHN</s3><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Jie Ding" sort="Jie Ding" uniqKey="Jie Ding" last="Jie Ding">JIE DING</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</s1><s2>Beijing, 100020</s2><s3>CHN</s3><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Chao-Yang Hospital Affiliated to Capital University of Medical Sciences</wicri:noRegion></affiliation></author><author><name sortKey="Jian Zhou" sort="Jian Zhou" uniqKey="Jian Zhou" last="Jian Zhou">JIAN ZHOU</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation></author><author><name sortKey="Hua Xiang" sort="Hua Xiang" uniqKey="Hua Xiang" last="Hua Xiang">HUA XIANG</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences</s1><s2>Beijing, 100080</s2><s3>CHN</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>République populaire de Chine</country><wicri:noRegion>Beijing, 100080</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">Applied microbiology and biotechnology</title><title level="j" type="abbreviated">Appl. microbiol. biotechnol.</title><idno type="ISSN">0175-7598</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Applied microbiology and biotechnology</title><title level="j" type="abbreviated">Appl. microbiol. biotechnol.</title><idno type="ISSN">0175-7598</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Coronavirus</term><term>Diagnosis</term><term>Escherichia coli</term><term>Gene expression</term><term>Lactococcus lactis</term><term>Nucleocapsid</term><term>Protein</term><term>Recombinant protein</term><term>Severe acute respiratory syndrome</term><term>Vaccination</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Nucléocapside</term><term>Protéine</term><term>Escherichia coli</term><term>Lactococcus lactis</term><term>Diagnostic</term><term>Coronavirus</term><term>Vaccination</term><term>Syndrome respiratoire aigu sévère</term><term>Expression génique</term><term>Protéine recombinante</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><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></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Huadong Pei" sort="Huadong Pei" uniqKey="Huadong Pei" last="Huadong Pei">HUADONG PEI</name></noRegion><name sortKey="Chaomin Sun" sort="Chaomin Sun" uniqKey="Chaomin Sun" last="Chaomin Sun">CHAOMIN SUN</name><name sortKey="Chaomin Sun" sort="Chaomin Sun" uniqKey="Chaomin Sun" last="Chaomin Sun">CHAOMIN SUN</name><name sortKey="Chen Wang" sort="Chen Wang" uniqKey="Chen Wang" last="Chen Wang">CHEN WANG</name><name sortKey="Hua Xiang" sort="Hua Xiang" uniqKey="Hua Xiang" last="Hua Xiang">HUA XIANG</name><name sortKey="Huadong Pei" sort="Huadong Pei" uniqKey="Huadong Pei" last="Huadong Pei">HUADONG PEI</name><name sortKey="Jian Zhou" sort="Jian Zhou" uniqKey="Jian Zhou" last="Jian Zhou">JIAN ZHOU</name><name sortKey="Jie Ding" sort="Jie Ding" uniqKey="Jie Ding" last="Jie Ding">JIE DING</name><name sortKey="Jingfang Liu" sort="Jingfang Liu" uniqKey="Jingfang Liu" last="Jingfang Liu">JINGFANG LIU</name><name sortKey="Jingfang Liu" sort="Jingfang Liu" uniqKey="Jingfang Liu" last="Jingfang Liu">JINGFANG LIU</name><name sortKey="Yueping Lu" sort="Yueping Lu" uniqKey="Yueping Lu" last="Yueping Lu">YUEPING LU</name><name sortKey="Yun Cheng" sort="Yun Cheng" uniqKey="Yun Cheng" last="Yun Cheng">YUN CHENG</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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