Interferon α 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-α. The expression of BoIFN-α in Escherichia coli was induced by addition of IPTG. Sequence analysis showed that the Chinese Luxi yellow cattle IFN-α gene is composed of 498 nucleotides, encoding a mature polypeptide of 166 amino acids. Compared with other BoIFN-α 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 40kD 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 5×105u/mg on MDBK/VSV cell system. The results showed that the expression plasmid was successfully constructed and BoIFN-α C2 protein was expressed in Escherichia coli. Moreover the purification had good effects on antiviral activities.
张永红,王长法,杨少华,高运东,王洪梅,李景鹏,仲跻峰. 重组鲁西黄牛α干扰素融合蛋白的表达及其抗病毒活性研究[J]. Chinese Journal of Biotechnology, 2007, 23(4):
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