Earticle

초록

영어

Peroxiredoxins are a group of thiol-specific antioxidant proteins that take six isoforms in vertebrates and allow the innate immune system to sense and detoxify reactive oxygen species. In this study, we identified and characterized the perxiredoxin-1 (SsPrdx1) cDNA sequence from the rockfish, Sebastes schlegelii. In silico analysis revealed that SsPrdx1 contained a 594 bp long open reading frame (ORF) encoding a protein of 198 amino acids, with a predicted molecular weight and theoretical isoelectric point of 21.97 kDa and 6.30, respectively. The SsPrdx1 gene comprised six exons linked by five introns, while peroxiredoxin signature motifs were found in the highly conserved third, fourth, and fifth exons. Phylogenetic analysis and sequence alignment suggested that SsPrdx1 is evolutionarily conserved and that its most closely related counterpart is Salarias fasciatus. Recombinant SsPrdx1 (rSsPrdx1) displayed supercoiled DNA protection and insulin disulfide reduction activities in a concentration-dependent manner, while cells transiently transfected with pcDNA3.1 (+)/SsPrdx1 exhibited significant cytoprotective effects under oxidative stress and wound healing activity. SsPrdx1 transcripts were constitutively expressed under normal physiological conditions, with the highest expression observed in the blood. Moreover, SsPrdx1 expression increased in the blood, spleen, and liver following immune provocation by LPS, poly I:C, and Streptococcus iniae injection. Thus, this study provides insights into the role of SsPrdx1 in rockfish immune protection.

목차

ABSTRACT
1. Introduction
2. Methodology
2.1. Animal husbandry
2.2. Preparation of black rockfish transcriptomic library
2.3. In silico SsPrdx1 characterization
2.4. Tissue sampling and immune stimulation
2.5. RNA extraction and cDNA synthesis
2.6. Relative quantification of SsPrdx1 expression
2.7. Fusion protein cloning, recombinant expression, and maltose affinitypurification
2.8. Functional assays
2.9. Statistical analysis
3. Results and discussion
3.1. Sequence characterization and structure prediction
3.2. Phylogenetic relationships, alignment analysis, and identity/similarity scores
3.3. SsPrdx1 mRNA expression under challenged and unchallengedconditions
3.4. Fusion protein overexpression and purification
3.5. Biological activity of SsPrdx1
4. Conclusion
CRediT authorship contribution statement
Acknowledgments
References

키워드

저자

• Rajamanthrilage Kasun Madusanka [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
• M.D. Neranjan Tharuka [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
• W.S.P. Madhuranga [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
• Seongdo Lee [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ]
• Jehee Lee [ Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, Jeju Self-Governing Province, 63243, Republic of Korea, Marine Science Institute, Jeju National University, Jeju Self-Governing Province, 63333, Republic of Korea ] Corresponding Author

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  해양과학연구소 연구논문집 [Bulletin of the Marine Science Institute]

• 간기

  폐간

• pISSN

  1225-5734

• 수록기간

  1977~2021

• 십진분류

  KDC 454 DDC 551