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Expression of bioactive porcine interferon-alpha in Lactobacillus casei

Author:
Ma, Shi-jie, Li, Kun, Li, Xin-Sheng, Guo, Xiao-Qing, Fu, Peng-Fei, Yang, Ming-Fan, Chen, Hong-Ying
Source:
World journal of microbiology & biotechnology 2014 v.30 no.9 pp. 2379-2386
ISSN:
0959-3993
Subject:
Lactobacillus brevis, Suid alphaherpesvirus 1, interferon-alpha, anaerobic conditions, lactose, messenger RNA, bioactive properties, secretion, reverse transcriptase polymerase chain reaction, Western blotting, enzyme-linked immunosorbent assay, swine, pH, Lactobacillus casei, virus replication, health services
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.
Agid:
803081