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《Chinese Journal of Veterinary Medicine》 2004-01
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Molecular cloning, expression and antiviral activity analysis of IFN-α gene of Holstein cow

WANG Ming,SHI Xi-ju,XIA Chun(College of Veterinary Medicine, China Agicultural University, Beijing 100094, China)  
Interferon-α gene was cloned from genomic DNA of Holstein cow by CPCR, and PCR product was digested with SphⅠ and HindⅢ,and then was ligated to PQE30 vector to construct recombinant expression plasmid PQE30/BoIFN-α.Sequence analysis showed that the cloned IFN-α gene Holstein cow was composed of 498 nucleotides, encoding 166 amino acids and only one point mutation was found when compared with BoIFN-α A subtype,the nucleotide and amino acid identity were 93.8%~(97.0%) and 90.4%~94.6% with published other subtypes, respectively. The constructed rBoIFN-α was expressed in E.coli JM109 by IPTG induction and the expression product was fusion protein with molecular weight of 20 kDa analyzed by SDS-PAGE and Western blot.Antiviral assays showed that the antiviral activities were 1.3~2.6×10~5 U/mg and 1.0~2.3×10~6 U/mg in CEF/VSV and MDBK/VSV cell lines,respectively.The rBoIFN-α protein could inhibit IBRV proliferation with 7.8×10~5 U/mg of antiproliferation activity in MDBK cell line. The rBoIFN-α is a potential biological agent for prevention and treatment of various kinds of bovine viral diseases.
【Fund】: 国家“十五”攻关项目 ( 2 0 0 2 BA5 14 A-16-4 )
【CateGory Index】: S852.4
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