Expression and purification of S5<inf>196-272</inf> and S6<inf>200-317</inf> proteins from Tilapia Lake Virus (TiLV) and their potential use as vaccines
Issued Date
2022-02-01
Resource Type
ISSN
10465928
eISSN
10960279
Scopus ID
2-s2.0-85118901313
Pubmed ID
34752859
Journal Title
Protein Expression and Purification
Volume
190
Rights Holder(s)
SCOPUS
Bibliographic Citation
Protein Expression and Purification Vol.190 (2022)
Suggested Citation
Lueangyangyuen A., Senapin S., Dong H.T., Unajak S., Wangkahart E., Khunrae P. Expression and purification of S5<inf>196-272</inf> and S6<inf>200-317</inf> proteins from Tilapia Lake Virus (TiLV) and their potential use as vaccines. Protein Expression and Purification Vol.190 (2022). doi:10.1016/j.pep.2021.106013 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/83852
Title
Expression and purification of S5<inf>196-272</inf> and S6<inf>200-317</inf> proteins from Tilapia Lake Virus (TiLV) and their potential use as vaccines
Other Contributor(s)
Abstract
Tilapia Lake Virus Disease (TiLVD) is caused by Tilapia Lake Virus (TiLV), and it has a cumulative mortality rate of up to 90% in Nile tilapia (Oreochromis niloticus). TiLV is a negative enveloped single-stranded RNA virus with 10 genomic segments. Segment 5 (S5) and segment 6 (S6) were predicted to include a signaling peptide, suggesting that the encoded proteins of these two segments may exist as part of the virus envelope. Based on bioinformatic predictions, the S5 and S6 proteins in this study were produced, including S527-343, S527-172, S5196-272, S630-317, S630-190, and S6200-317. All proteins were tested for their expression in Escherichia coli. Only S5196-272 and S6200-317 were expressed as soluble and insoluble proteins, respectively. The soluble protein was purified using affinity chromatography, whereas the insoluble protein was solubilized using 6 M urea lysis buffer before purification. Both proteins were further purified using gel filtration chromatography, and the results showed a symmetric peak of both proteins suggested a high degree of uniformity in the conformation of these proteins. Antigenicity results indicated that these proteins were recognized by serum from TiLV-infected fish. The immunization tests revealed that serum antibodies levels in Nile tilapia produced by S5196-272 and S6200-317 were significantly increased (p-value < 0.05) at 7 days post-immunization (dpi) compared to antibody levels on Day 0 (D0). All the results combined suggested a potential vaccine candidate of S5 and S6 for TiLV protection in Nile tilapia.