title: Expression of bioactive porcine interferon-alpha in Lactobacillus casei.
abstract: In this study, we constructed an expression cassette containing the inducible lac promoter and the secretion signal from an S-layer protein of Lactobacillus brevis for the expression of porcine interferon-alpha (IFN-α) in Lactobacillus casei (Lb. casei). Reverse-transcriptase PCR verified the presence of porcine IFN-α mRNA in the recombinant Lb. casei. The porcine IFN-α protein expressed in the recombinant Lb. casei was identified by both Western blot analysis and ELISA. We used various pH values and induction times to optimize the yield of IFN-α, and found that induction with 0.8% lactose for 16 h under anaerobic conditions produced the highest concentrations of IFN-α. Furthermore, the activity of porcine IFN-α in the cultural supernatant was evaluated on ST cells infected with pseudorabies virus. The results revealed that porcine IFN-α inhibited virus replication in vitro. The findings of our study indicate that recombinant Lb. casei producing porcine IFN-α has great potential for use as a novel oral antiviral agent in animal healthcare.
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Nongye Road 63#, Zhengzhou, 450002, Henan Province, People's Republic of China.
authors.forename: Shi-jie
authors.initials: SJ
authors.lastname: Ma
authors.forename: Kun
authors.initials: K
authors.lastname: Li
authors.forename: Xin-Sheng
authors.initials: XS
authors.lastname: Li
authors.forename: Xiao-Qing
authors.initials: XQ
authors.lastname: Guo
authors.forename: Peng-Fei
authors.initials: PF
authors.lastname: Fu
authors.forename: Ming-Fan
authors.initials: MF
authors.lastname: Yang
authors.forename: Hong-Ying
authors.initials: HY
authors.lastname: Chen
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D011558:Herpesvirus 1, Suid / Q000187:drug effects / Q000502:physiology; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D007780:Lacticaseibacillus casei / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D015533:Transcriptional Activation; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D016898:Interferon-alpha; D011994:Recombinant Proteins