Fads2b Plays a Dominant Role in ∆6/∆5 Desaturation Activities Compared with Fads2a in Common Carp (Cyprinus carpio)
Abstract
:1. Introduction
2. Results
2.1. The Differences in Gene and Protein Structures of Common Carp fads2a and fads2b
2.2. Divergent Spatial and Temporal Expression Patterns of fads2a and fads2b In Vivo
2.3. Differences in Desaturation Activities of Common Carp Fads2a and Fads2b in Transgenic Yeast
2.4. Molecular Docking Analyses of Fads2a and Fads2b
2.5. Promoter Activity Analysis of fads2a and fads2b
3. Discussion
4. Materials and Methods
4.1. Animals and Ethics Statement
4.2. Sequence and Structure Analysis of Two Common Carp fads2 Genes
4.3. Phylogenetic Analysis
4.4. GC–MS Analysis of Common Carp Embryos at Different Developmental Stages
4.5. Heterologous Expression of fads2 ORFs in Yeast
4.6. Expression of fads2 Genes during Common Carp Ontogeny
4.7. Whole-Mount In Situ Hybridization (WISH)
4.8. Molecular Docking
4.9. Cloning of fads2a/fads2b Promoters and Construction of Expression Plasmids
4.10. Cell Culture, Transfection and Luciferase Assay
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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FA Substrate | Product | FA Substrate Peak Area (pA·s) | Product Peak Area (pA·s) | Conversion (%) | |
---|---|---|---|---|---|
Fads2a | C18:2n-6 | C18:3n-6 | 30,945.8 | 101.4 | 0.33 |
Fads2a | C18:3n-3 | C18:4n-3 | 325.5 | 4.8 | 1.45 |
Fads2a | C20:3n-6 | C20:4n-6 | 3817.5 | 10.9 | 0.28 |
Fads2a | C20:4n-3 | C20:5n-3 | 2194.9 | 17.2 | 0.78 |
Fads2b | C18:2n-6 | C18:3n-6 | 8603.6 | 51.4 | 0.59 |
Fads2b | C18:3n-3 | C18:4n-3 | 7850.4 | 318.2 | 3.89 |
Fads2b | C20:3n-6 | C20:4n-6 | 2762.5 | 8.9 | 0.32 |
Fads2b | C20:4n-3 | C20:5n-3 | 3870 | 42 | 1.07 |
Fatty Acids | Docking Energy (kcal/mol) | |
---|---|---|
Fads2a | Fads2b | |
C18:2n-6 | −5.86 | −5.04 |
C18:3n-3 | −5.83 | −6.13 |
C20:3n-6 | −5.98 | −5.54 |
C20:4n-3 | −5.73 | −5.84 |
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Zhao, R.; Yang, C.-R.; Wang, Y.-X.; Xu, Z.-M.; Li, S.-Q.; Li, J.-C.; Sun, X.-Q.; Wang, H.-W.; Wang, Q.; Zhang, Y.; et al. Fads2b Plays a Dominant Role in ∆6/∆5 Desaturation Activities Compared with Fads2a in Common Carp (Cyprinus carpio). Int. J. Mol. Sci. 2023, 24, 10638. https://doi.org/10.3390/ijms241310638
Zhao R, Yang C-R, Wang Y-X, Xu Z-M, Li S-Q, Li J-C, Sun X-Q, Wang H-W, Wang Q, Zhang Y, et al. Fads2b Plays a Dominant Role in ∆6/∆5 Desaturation Activities Compared with Fads2a in Common Carp (Cyprinus carpio). International Journal of Molecular Sciences. 2023; 24(13):10638. https://doi.org/10.3390/ijms241310638
Chicago/Turabian StyleZhao, Ran, Chen-Ru Yang, Ya-Xin Wang, Zi-Ming Xu, Shang-Qi Li, Jin-Cheng Li, Xiao-Qing Sun, Hong-Wei Wang, Qi Wang, Yan Zhang, and et al. 2023. "Fads2b Plays a Dominant Role in ∆6/∆5 Desaturation Activities Compared with Fads2a in Common Carp (Cyprinus carpio)" International Journal of Molecular Sciences 24, no. 13: 10638. https://doi.org/10.3390/ijms241310638