+ 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

Expression and purification of recombinant SARS coronavirus spike protein

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

A novel coronavirus (SARS-coronavirus, SARS-CoV) was discovered in association with cases of severe acute respiratory syndrome (SARS) recently. The first step in coronavirus infection is binding of the viral spike protein to certain receptor on host cells. The spike protein is the main surface antigen of the coronavirus and there should be antibodies against spike protein in patients' serum. Thus, to develop and expression protein fragment from spike protein gene are the purposes of this experiment. Partial spike gene fragments (751apprx1925 bp, 2005apprx3410 bp, 1apprx1925 bp and 32apprx3659 bp) and its intact gene were cloned into pET32 or pGEX vectors, and transformed into competent Escherichia coli BL21 (DE3) (pLysS), respectively. 63, 78, 98, 160 and 164 kD fusion proteins were successfully expressed with amounts of 35%, 34%, 24%, 17% and 5% of total cell protein. The soluble parts of the cell crude extract were then partially purified by GST affinity chromatography.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 010636502

Download citation: RISBibTeXText

PMID: 12897976

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

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 recombinant spike protein of SARS-coronavirus in vaccinia virus and analysis of its immunogenicity. Bing du Xue Bao 23(4): 287-291, 2007

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

Expression, purification and identification of recombinant SARS coronavirus membrane protein. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao Acta Biochimica et Biophysica Sinica 35(12): 1140-1144, 2003

Antigenicity and receptor-binding ability of recombinant SARS coronavirus spike protein. Biochemical and Biophysical Research Communications 313(4): 938-947, 2004

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

Cloning, sequencing, expression, and purification of SARS-associated coronavirus nucleocapsid protein for serodiagnosis of SARS. Journal of Clinical Virology 30(4): 309-312, 2004

Intranasal vaccination of recombinant adeno-associated virus encoding receptor-binding domain of severe acute respiratory syndrome coronavirus (SARS-CoV) spike protein induces strong mucosal immune responses and provides long-term protection against SARS-CoV infection. Journal of Immunology 180(2): 948-956, 2008

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

Mouse studies of SARS coronavirus-specific immune responses to recombinant replication-defective adenovirus expressing SARS coronavirus N protein. Hong Kong Medical Journal 15(Suppl. 2): 33-36, 2009

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

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, 2005