+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Prokaryotic expression of SARS coronavirus spike protein and construction of its DNA vaccine

Prokaryotic expression of SARS coronavirus spike protein and construction of its DNA vaccine

Di 1 Jun Yi Da Xue Xue Bao 25(1): 33-36

To study the immunological characteristics of the spike (S) protein of SARS coronavirus (SARS-CoV) and analyze the feasibility of using this protein as the component for SARS vaccine development. The two truncated fragments of S gene were separately cloned into the prokaryotic expression vector pET-15b and expressed in E.coli. The resulting recombinant proteins, rS(a) and rS(b), were purified by affinity chromatography. The full-length S gene was cloned into the eukaryotic expression plasmid pSecTagB to prepare recombinant plasmid pSecS as the DNA vaccine to immunize BALB/c mice for inducing the secretion of anti-SARS-CoV protein. The immunological effect of anti-SARS-CoV antibody was tested with purified rS(a) and rS(b) proteins by enzyme-linked immunosorbent assay (ELISA). Both the truncated recombinant proteins were expressed in soluble forms and reacted specifically with the sera from immunized pSecS mice and clinically diagnosed SARS patients. The prokaryotically expressed recombinant truncated S protein had similar antigenicity with SARS-CoV S protein. The recombinant protein could be used as an antigen for detecting the serum of SARS CoV-infected patients. The SARS-CoV S gene vaccine could induce the production of specific antibody, which offers clues for the research of SARS DNA vaccine.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 050041573

Download citation: RISBibTeXText

PMID: 15683993

Related references

Prokaryotic expression, refolding, and purification of fragment 450-650 of the spike protein of SARS-coronavirus. Protein Expression and Purification 39(2): 169-174, 2005

Prokaryotic expression of S2 extracellular domain of SARS coronavirus spike protein and its fusion with Hela cell membrane. Nan Fang Yi Ke Da Xue Xue Bao 29(3): 381-386, 2009

SARS coronavirus spike polypeptide DNA vaccine priming with recombinant spike polypeptide from Escherichia coli as booster induces high titer of neutralizing antibody against SARS coronavirus. Vaccine 23(42): 4959-4968, 2005

Expression of SARS coronavirus membrane protein and its DNA vaccine construction. Virologica Sinica 20(2): 113-116, 2005

Expression of SARS coronavirus nucleocapsid protein and construction of its DNA vaccine. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 21(6): 731, 2005

Differential sensitivities of severe acute respiratory syndrome (SARS) coronavirus spike polypeptide enzyme-linked immunosorbent assay (ELISA) and SARS coronavirus nucleocapsid protein ELISA for serodiagnosis of SARS coronavirus pneumonia. Journal of Clinical Microbiology 43(7): 3054-3058, 2005

Prokaryotic expression of SARS coronavirus nonstructural protein 9. Virologica Sinica 20(5): 552-554, 2005

Heterologous viral RNA export elements improve expression of severe acute respiratory syndrome (SARS) coronavirus spike protein and protective efficacy of DNA vaccines against SARS. Virology 363(2): 288-302, 2007

Development of a SARS Coronavirus Vaccine from Recombinant Spike Protein Plus Delta Inulin Adjuvant. Methods in Molecular Biology 1403: 269-284, 2016

Immunogenicity of a receptor-binding domain of SARS coronavirus spike protein in mice: implications for a subunit vaccine. Vaccine 25(1): 136-143, 2007

Cloning and expression of SARS Coronavirus spike protein fragment 1. Chinese Journal of Zoonoses 19(5): 13-16, 2003

Expression and purification of recombinant SARS coronavirus spike protein. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao Acta Biochimica et Biophysica Sinica 35(8): 774-778, 2003

Synthetic peptide studies on the severe acute respiratory syndrome (SARS) coronavirus spike glycoprotein: perspective for SARS vaccine development. Clinical Chemistry 50(6): 1036-1042, 2004

Immunogenicity and protection efficacy of monomeric and trimeric recombinant SARS coronavirus spike protein subunit vaccine candidates. Viral Immunology 26(2): 126-132, 2013