title: Prokaryotic-expressed porcine IFNα1-THYα1 fusion proteins exert IFN and THY activities in vitro.
abstract: Combined use of interferon (IFN) and thymosin (THY) holds a stronger antiviral effect than when applied individually because of their coordination and complementary action. In this study, prokaryotic expressed porcine IFNα1 (poIFNα1) or the porcine IFNα1-THYα1 fusion protein coding with the Escherichia coli preferred codon sequences connected by the three different linkers were gained in the unlabeled pRSFDDuet-1 expression systems and purified using the strong anion-exchange chromatography and hydrophobic chromatography (among which, one was digested by thrombin because the cleavage site was included in the linker). Then, the activities of IFN and THY in the fusion protein were detected using the cytopathic effect inhibition assay and T-cell activity assays. SDS PAGE and western blotting results showed that the poIFNα1 or the three poIFNα1-THYα1 fusion proteins with three different linkers were expressed solubly in E. coli. The poIFNα1 protein and three types of poIFNα1-THYα1 fusion proteins with >90% purity were gained. The poIFNα1-LinkerB-THYα1 fusion protein showed the highest interferon activity compared with the others (P < 0.001), and the poIFNα1-LinkerA-THYα1 fusion protein highest thymosin activity (P < 0.05). In this study, a preliminary experiment was conducted for the expression of the poIFNα1 and THYα1 fusion proteins.
authors.affiliation: College of Animal Science, Jilin University, Changchun 130062, China; China Institute of Veterinary Drug Control, Beijing 100081, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Wang
authors.affiliation: College of Animal Science, Jilin University, Changchun 130062, China; Five-Star Animal Health Pharmaceutical Factory of Jilin Province, Changchun 130062, China.
authors.forename: Songcai
authors.initials: S
authors.lastname: Liu
authors.affiliation: College of Animal Science, Jilin University, Changchun 130062, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Shi
authors.affiliation: College of Animal Science, Jilin University, Changchun 130062, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.affiliation: College of Animal Science, Jilin University, Changchun 130062, China.
authors.forename: Yunyun
authors.initials: Y
authors.lastname: Cheng
authors.affiliation: College of Animal Science, Jilin University, Changchun 130062, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Su
authors.affiliation: College of Animal Science, Jilin University, Changchun 130062, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Lu
authors.affiliation: College of Animal Science, Jilin University, Changchun 130062, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Yu
authors.affiliation: College of Animal Science, Jilin University, Changchun 130062, China. Electronic address: haolinlin@jlu.edu.cn.
authors.forename: Linlin
authors.initials: L
authors.lastname: Hao
mesh_terms: D000818:Animals; D066298:In Vitro Techniques; D016898:Interferon-alpha / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D013552:Swine; D013917:Thrombin / Q000378:metabolism; D000077596:Thymalfasin; D013947:Thymosin / Q000031:analogs & derivatives* / Q000235:genetics / Q000502:physiology
chemical_list: D016898:Interferon-alpha; D011993:Recombinant Fusion Proteins; D013947:Thymosin; D013917:Thrombin; D000077596:Thymalfasin
keywords: Fusion protein; Porcine IFN-α1; Purification; THY-α1