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
in Zh
| WPRIM
| ID: wpr-327956
Responsible library:
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.
Full text:
1
Database:
WPRIM
Main subject:
Antiviral Agents
/
Recombinant Fusion Proteins
/
Molecular Sequence Data
/
Base Sequence
/
Amino Acid Sequence
/
Interferon-alpha
/
Sequence Analysis
/
Rotavirus
/
Escherichia coli
/
Genetics
Limits:
Animals
Language:
Zh
Journal:
Chinese Journal of Biotechnology
Year:
2007
Document type:
Article