Cloning, purification, and antigenic characterization of three recombinant fragments derived from SARS-CoV S1 domain / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 275-278, 2005.
Artículo
en Chino
| WPRIM
| ID: wpr-333023
ABSTRACT
<p><b>OBJECTIVE</b>The present study aimed to clone and express three fragments of genomic RNA derived from SARS associated coronavirus (SARS-CoV) S1 domain and to study its immunogenicity.</p><p><b>METHODS</b>The S1 domain gene was amplified by PCR with specific primers and was inserted into the prokaryotic expression vector pQE-30. Three fragments (40-751, 746-1344 and 746-2001 bp) derived from S1 domain produced after the recombinant plasmid (pQE-30/S1) was digested by restriction endonucleases. The three fragments were cloned into pQE-30 and expressed in M15 strains of Escherichia coli. The expression products, designated S1a, S1b and S1c respectively, were purified by Ni affinity chromatography. The immunogenicity was analyzed by Western Blot and ELISA using serologically confirmed sera from SARS patients and the sera from healthy donors was used as control at the same assay.</p><p><b>RESULTS</b>Three recombinant plasmids (pQE-30/S1a, pQE-30/S1b, pQE-30/S1c) were constructed.Fusion proteins with relative molecular mass of 26,700, 22,500 and 46,000 dalton were successfully expressed with amounts of 35%, 35% and 30% of total cell protein and purified by Ni affinity chromatography, respectively. Western Blot and ELISA analysis showed that the S1c protein could be specifically recognized by the sera from SARS patients.</p><p><b>CONCLUSION</b>The recombinant S1c protein was a good immunogen and has the potential to be used as a vaccine against SARS-CoV infection.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Plásmidos
/
Virología
/
Sangre
/
Proteínas Recombinantes
/
Inmunoglobulina G
/
Inmunoglobulina M
/
Ensayo de Inmunoadsorción Enzimática
/
Proteínas del Envoltorio Viral
/
Western Blotting
/
Reacción en Cadena de la Polimerasa
Límite:
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Experimental and Clinical Virology
Año:
2005
Tipo del documento:
Artículo
Similares
MEDLINE
...
LILACS
LIS