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Chinese Journal of Biotechnology ; (12): 534-539, 2005.
Artículo en Chino | WPRIM | ID: wpr-305207

RESUMEN

To construct plasmid of recombinant protein candidate vaccine of respiratory syncytial virus, express it in E. coli, and to investigate its immunogenicity and protective efficacy. A CD8+ T cell epitope from respiratory syncytial virus (RSV) M2 protein F/M2:81 - 95 and the G:125-225 (G1) gene fragments from RSV-G protein containing B cell epitopes were amplified by PCR method and then inserted into the prokaryotic expression vector pET-DsbA after bonding to a linker. The fusion protein DsbA-G1-Linker-F/M2:81-95 (D-G1LF/M2) was expressed successfully in E. coli BL21 (DE3). The product was proved to be RSV-specific by Western-blot. After purified by affinity chromatography on Ni+ Sepharose and renatured by gradient dialysis. D-G1LF/M2 was used to immune BALB/c mice. D-G1LF/M2 induced high anti-D-G1LF/M2 IgG, anti-RSV IgG and neutralizing antibody titers in serum and lung of BALB/c mice, and elicied RSV-specific CTL responses. The IgG subclass distribution revealed that IgG1/IgG2a ratio was 2.66. Viral titration indicated that D-G1LF/M2 could protect BALB/c mice against RSV challenge in lung.


Asunto(s)
Animales , Humanos , Ratones , Anticuerpos Antivirales , Sangre , Alergia e Inmunología , Escherichia coli , Genética , Metabolismo , Inmunoglobulina G , Sangre , Alergia e Inmunología , Ratones Endogámicos BALB C , Plásmidos , Genética , Alergia e Inmunología , Proteínas Recombinantes de Fusión , Genética , Alergia e Inmunología , Infecciones por Virus Sincitial Respiratorio , Vacunas contra Virus Sincitial Respiratorio , Genética , Alergia e Inmunología , Virus Sincitial Respiratorio Humano , Genética , Alergia e Inmunología , Proteínas del Envoltorio Viral , Genética , Proteínas Virales de Fusión , Genética , Proteínas Virales , Genética
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