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Secretory Production of Functional Grouper Type I Interferon from Epinephelus septemfasciatus in Escherichia coli and Bacillus subtilis
- Source :
- International Journal of Molecular Sciences, Vol 21, Iss 4, p 1465 (2020), International Journal of Molecular Sciences, Volume 21, Issue 4
- Publication Year :
- 2020
- Publisher :
- MDPI AG, 2020.
-
Abstract
- Nervous necrosis virus (NNV) results in high mortality rates of infected marine fish worldwide. Interferons (IFNs) are cytokines in vertebrates that suppress viral replication and regulate immune responses. Heterologous overexpression of fish IFN in bacteria could be problematic because of protein solubility and loss of function due to protein misfolding. In this study, a protein model of the IFN-&alpha<br />of Epinephelus septemfasciatus was built based on comparative modeling. In addition, PelB and SacB signal peptides were fused to the N-terminus of E. septemfasciatus IFN-&alpha<br />for overexpression of soluble, secreted IFN in Escherichia coli (E-IFN) and Bacillus subtilis (B-IFN). Cytotoxicity tests indicated that neither recombinant grouper IFN-&alpha<br />were cytotoxic to a grouper head kidney cell line (GK). The GK cells stimulated with E-IFN and B-IFN exhibited elevated expression of antiviral Mx genes when compared with the control group. The NNV challenge experiments demonstrated that GK cells pretreated or co-treated with E-IFN and B-IFN individually had three times the cell survival rates of untreated cells, indicating the cytoprotective ability of our recombinant IFNs. These data provide a protocol for the production of soluble, secreted, and functional grouper IFN of high purity, which may be applied to aquaculture fisheries for antiviral infection.
- Subjects :
- 0301 basic medicine
nervous necrosis virus
Heterologous
Bacillus subtilis
epinephelus septemfasciatus
Biology
medicine.disease_cause
Catalysis
Microbiology
Inorganic Chemistry
lcsh:Chemistry
03 medical and health sciences
Immune system
Interferon
medicine
Cytotoxic T cell
Grouper
signal peptide
Physical and Theoretical Chemistry
Molecular Biology
Escherichia coli
lcsh:QH301-705.5
Spectroscopy
030102 biochemistry & molecular biology
Organic Chemistry
General Medicine
interferon
biology.organism_classification
Computer Science Applications
030104 developmental biology
aquaculture
lcsh:Biology (General)
lcsh:QD1-999
Cell culture
medicine.drug
Subjects
Details
- Language :
- English
- ISSN :
- 14220067
- Volume :
- 21
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- International Journal of Molecular Sciences
- Accession number :
- edsair.doi.dedup.....afe929c0d0613fdbea8a88f66ec4743b