Expression, purification of recombinant Luxi yellow cattle IFN-alpha fusion protein and its antiviral activities / 生物工程学报
Chinese Journal of Biotechnology
; (12): 730-734, 2007.
Article
en Zh
| WPRIM
| ID: wpr-327956
Biblioteca responsable:
WPRO
ABSTRACT
Interferon a gene was cloned from genomic DNA of Chinese Luxi yellow cattle by PCR, and the PCR product was inserted into vector pET32a( + ) to make a recombinant plasmid pET32a( + )/BoIFN-alpha. The expression of BoIFN-alpha in Escherichia coli was induced by addition of IPTG. Sequence analysis showed that the Chinese Luxi yellow cattle IFN-alpha gene is composed of 498 nucleotides, encoding a mature polypeptide of 166 amino acids. Compared with other BoIFN-alpha subtypes, it shares the highest identity of 97.6% to the C-subtype. SDS-PAGE results showed that recombinant proteins were expressed in inclusion bodies in Escherichia coli with molecular weight of 40 kD and the recombinant proteins accounted for 26.7% of the whole proteins.The expressed product was purified by affinity chromatography with immobilized nickel chelating NTA (Ni-NTA) and its antiviral activities were tested on MDBK/VSV cell system. Its antiviral activities were 5 x 10(5) u/mg on MDBK/VSV cell system. The results showed that the expression plasmid was successfully constructed and BoIFN-alpha C2 protein was expressed in Escherichia coli. Moreover the purification had good effects on antiviral activities.
Texto completo:
1
Índice:
WPRIM
Asunto principal:
Antivirales
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Proteínas Recombinantes de Fusión
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Datos de Secuencia Molecular
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Secuencia de Bases
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Secuencia de Aminoácidos
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Interferón-alfa
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Análisis de Secuencia
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Rotavirus
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Escherichia coli
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Genética
Límite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2007
Tipo del documento:
Article