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.
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This work was funded by the Important Henan Province Science & Technology Specific Projects (111100110300), and the Innovation Scientists and Technicians Troop Construction Projects of Zhengzhou City (10CXTD148).
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Shi-jie Ma and Kun Li have contributed equally to this work.
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Ma, Sj., Li, K., Li, XS. et al. Expression of bioactive porcine interferon-alpha in Lactobacillus casei . World J Microbiol Biotechnol 30, 2379–2386 (2014). https://doi.org/10.1007/s11274-014-1663-7
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DOI: https://doi.org/10.1007/s11274-014-1663-7