The invention belongs to the field of interferons, and particularly relates to a high-activity porcine
interferon alpha
mutant with 7
mutation loci and a preparing method and application thereof. According to the high-activity porcine
interferon alpha
mutant with the 7
mutation loci and the preparing method and application thereof, a porcine
interferon alpha gene is modified to obtain a new
gene sequence, and through modification, the
biological activity of the interferon is improved. An overlapped
PCR method is applied to conducting site-
specific mutation on the natural porcine
interferon alpha gene, the modified
gene is cloned to a pET-32a carrier to construct a recombinant
plasmid, then the recombinant
plasmid which is accurately identified is cloned into Rosetta expression
bacteria forinduction expression and identification, recombinant
protein is expressed in the form of an inclusion body, the molecular
mass of the
protein is about 32 KDa, after purification and
dialysis treatment, the porcine
interferon alpha mutant with high purity is obtained, and the antiviral activity is determined through
cytopathic effect inhibition. As is shown through results, the antiviral activityof the
mutant protein obtained through experiments is 8.15*10<8> U / mg, and increased by about 200 times compared with that of wild-type interferons expressed
in vitro.