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[Molecular cloning and expression of the severe acute respiratory syndrome-associated coronavirus nucleocapsid protein and its clinical application].
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2005 Mar; 19(1):64-7.ZS

Abstract

OBJECTIVE

To clone and express nucleocapsid (N) protein of the severe acute respiratory syndrome (SARS)-associated coronavirus, and to evaluate its antigenicity and application value in the development of serological diagnostic test for SARS.

METHODS

SARS-associated coronavirus N protein gene was amplified from its genomic RNA by reverse transcript nested polymerase chain reaction (RT-nested-PCR) and cloned into pBAD/Thio-TOPO prokaryotic expression vector. The recombinant N fusion protein was expressed and purified, and its antigenicity and specificity was analyzed by Western Blot, to establish the recombinant N protein-based ELISA for detection of IgG antibodies to SARS-associated coronavirus, and SARS-associated coronavirus lysates-based ELISA was compared parallelly.

RESULTS

The recombinant expression vector produced high level of the N fusion protein after induction, and that protein was purified successfully by affinity chromatography and displayed higher antigenicity and specificity as compared with whole virus lysates.

CONCLUSION

The recombinant SARS-associated coronavirus N protein possessed better antigenicity and specificity and could be employed to establish a new, sensitive, and specific ELISA for SARS diagnosis.

Authors+Show Affiliations

Lu J
Shenzhen Donghu Hospital, Shenzhen, China.
Zhou BP
No affiliation info available
Zhou YS
No affiliation info available
Jiang XL
No affiliation info available
Wen LX
No affiliation info available
Le XH
No affiliation info available
Li B
No affiliation info available
Xu LM
No affiliation info available
Li LX
No affiliation info available

MeSH

Antibodies, ViralBlotting, WesternCloning, MolecularCoronavirus Nucleocapsid ProteinsEnzyme-Linked Immunosorbent AssayGene ExpressionGenome, ViralHumansImmunoglobulin GNucleocapsid ProteinsRNA, ViralRecombinant ProteinsReverse Transcriptase Polymerase Chain ReactionSevere acute respiratory syndrome-related coronavirusSevere Acute Respiratory Syndrome

Pub Type(s)

Journal Article

Language

chi

PubMed ID

16201478

Citation

Lu, Jian, et al. "[Molecular Cloning and Expression of the Severe Acute Respiratory Syndrome-associated Coronavirus Nucleocapsid Protein and Its Clinical Application]." Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi = Zhonghua Shiyan He Linchuang Bingduxue Zazhi = Chinese Journal of Experimental and Clinical Virology, vol. 19, no. 1, 2005, pp. 64-7.
Lu J, Zhou BP, Zhou YS, et al. [Molecular cloning and expression of the severe acute respiratory syndrome-associated coronavirus nucleocapsid protein and its clinical application]. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2005;19(1):64-7.
Lu, J., Zhou, B. P., Zhou, Y. S., Jiang, X. L., Wen, L. X., Le, X. H., Li, B., Xu, L. M., & Li, L. X. (2005). [Molecular cloning and expression of the severe acute respiratory syndrome-associated coronavirus nucleocapsid protein and its clinical application]. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi = Zhonghua Shiyan He Linchuang Bingduxue Zazhi = Chinese Journal of Experimental and Clinical Virology, 19(1), 64-7.
Lu J, et al. [Molecular Cloning and Expression of the Severe Acute Respiratory Syndrome-associated Coronavirus Nucleocapsid Protein and Its Clinical Application]. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2005;19(1):64-7. PubMed PMID: 16201478.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Molecular cloning and expression of the severe acute respiratory syndrome-associated coronavirus nucleocapsid protein and its clinical application]. AU - Lu,Jian, AU - Zhou,Bai-ping, AU - Zhou,Yu-sen, AU - Jiang,Xiao-ling, AU - Wen,Li-xia, AU - Le,Xiao-hua, AU - Li,Bing, AU - Xu,Liu-mei, AU - Li,Li-xiong, PY - 2005/10/6/pubmed PY - 2009/1/6/medline PY - 2005/10/6/entrez SP - 64 EP - 7 JF - Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology JO - Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi VL - 19 IS - 1 N2 - OBJECTIVE: To clone and express nucleocapsid (N) protein of the severe acute respiratory syndrome (SARS)-associated coronavirus, and to evaluate its antigenicity and application value in the development of serological diagnostic test for SARS. METHODS: SARS-associated coronavirus N protein gene was amplified from its genomic RNA by reverse transcript nested polymerase chain reaction (RT-nested-PCR) and cloned into pBAD/Thio-TOPO prokaryotic expression vector. The recombinant N fusion protein was expressed and purified, and its antigenicity and specificity was analyzed by Western Blot, to establish the recombinant N protein-based ELISA for detection of IgG antibodies to SARS-associated coronavirus, and SARS-associated coronavirus lysates-based ELISA was compared parallelly. RESULTS: The recombinant expression vector produced high level of the N fusion protein after induction, and that protein was purified successfully by affinity chromatography and displayed higher antigenicity and specificity as compared with whole virus lysates. CONCLUSION: The recombinant SARS-associated coronavirus N protein possessed better antigenicity and specificity and could be employed to establish a new, sensitive, and specific ELISA for SARS diagnosis. SN - 1003-9279 UR - https://www.unboundmedicine.com/medline/citation/16201478/[Molecular_cloning_and_expression_of_the_severe_acute_respiratory_syndrome_associated_coronavirus_nucleocapsid_protein_and_its_clinical_application]_ DB - PRIME DP - Unbound Medicine ER -
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