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Molecular characterization and expression of CD48 in Nile tilapia (Oreochromis niloticus) in response to different stimulus.
- Source :
-
Fish & shellfish immunology [Fish Shellfish Immunol] 2020 Feb; Vol. 97, pp. 515-522. Date of Electronic Publication: 2019 Dec 14. - Publication Year :
- 2020
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Abstract
- CD48 is a glycosylphosphatidylinositol-anchored protein involved in lymphocyte adhesion, activation, and costimulation. In this study, the CD48 gene of tilapia (Oreochromis niloticus, named On-CD48), was cloned from the head kidney of tilapia. The coding sequences is 654 bp and encoding 217 amino acids. The deduced amino acid sequence of On-CD48 with an estimated molecular weight of 24.4 kDa and a theoretical pI of 5.03. Amino acid alignment indicated that it had two immunoglobulin-like domain conserved region. In healthy tilapia, the On-CD48 could be detected in all the examined tissues and the highest expression level in the spleen. The expression of On-CD48 in the spleen and head kidney was decreased after immunized by formalin-inactivated Streptococcus agalactiae, and the peak was observed in the spleen at 24 h and appeared again at 96 h, and in the head kidney gradual decline before 48 h then gradually increased to the original level. qPCR analysis of inactivated S. agalactiae, LPS and Poly I:C stimulated at the whole lymphocyte level showed that the stimulation of the Poly I:C was more sensitive. Prokaryotic expression results showed that efficient expression of On-CD48 protein could be realized after induced with 0.5 mmol L <superscript>-1</superscript> IPTG in E. coli BL21 (DE3) for 10 h at 18 °C. The result of subcellular localization showed that On-CD48 were evenly distributed in the whole cell of HEK-293T. Western Blot confirmed that the molecular weight of the recombinant On-CD48 was about 21 kDa, consistent with the predicted result. The results of this study will lay a strong foundation for the further study of On-CD48 molecular function in tilapia.<br />Competing Interests: Declaration of competing interest The authors declare no conflict of interest.<br /> (Copyright © 2019 Elsevier Ltd. All rights reserved.)
- Subjects :
- Animals
CD48 Antigen immunology
Cichlids genetics
Fish Diseases immunology
Fish Diseases microbiology
Fish Proteins immunology
Gene Expression Regulation
HEK293 Cells
Humans
Lipopolysaccharides pharmacology
Lymphocytes drug effects
Phylogeny
Poly I-C pharmacology
Streptococcal Infections immunology
CD48 Antigen genetics
Cichlids immunology
Fish Proteins genetics
Streptococcal Infections veterinary
Subjects
Details
- Language :
- English
- ISSN :
- 1095-9947
- Volume :
- 97
- Database :
- MEDLINE
- Journal :
- Fish & shellfish immunology
- Publication Type :
- Academic Journal
- Accession number :
- 31846775
- Full Text :
- https://doi.org/10.1016/j.fsi.2019.12.034