Identification and Functional Analysis of Ras-Related Associated with Diabetes Gene (rrad) in Edwardsiella piscicida-Resistant Individuals of Japanese Flounder (Paralichthys olivaceus)
Abstract
:1. Introduction
2. Results
2.1. Sequence and Characteristics of rrad in P. olivaceus
2.2. Tissue Expression Distribution of rrad
2.3. Expression of rrad in Immune-Related Tissue after E. piscicida Infection
2.4. In Vitro Stimulation with PAMPs
2.5. Methylation Analysis of rrad in the Spleen
2.6. Functional Characterization of rrad Promoter
2.7. Luciferase Activity and Flow Cytometry Analyses
2.8. Protein–Protein Interaction (PPI) Network of rrad
2.9. Effect of the Knockdown of rrad and Other Related Genes via siRNA Transfection
3. Discussion
4. Materials and Methods
4.1. Fish Sample Preparation
4.2. Gene Cloning and cDNA Synthesis
4.3. Bioinformatic Analysis
4.4. Quantitative Real-Time PCR (qPCR) Analysis
4.5. Effect of rrad on the Survival Integrity of Target Bacteria
4.6. DNA Methylation Status of rrad in Different Individuals
4.7. rrad Promoter Activity Analysis
4.8. SiRNA Design and rrad Knockdown in Gill Cell Line of P. olivaceus
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Zhu, Y.; Yang, X.; Yang, Y.; Yan, X.; Li, C.; Chen, S. Identification and Functional Analysis of Ras-Related Associated with Diabetes Gene (rrad) in Edwardsiella piscicida-Resistant Individuals of Japanese Flounder (Paralichthys olivaceus). Int. J. Mol. Sci. 2024, 25, 10532. https://doi.org/10.3390/ijms251910532
Zhu Y, Yang X, Yang Y, Yan X, Li C, Chen S. Identification and Functional Analysis of Ras-Related Associated with Diabetes Gene (rrad) in Edwardsiella piscicida-Resistant Individuals of Japanese Flounder (Paralichthys olivaceus). International Journal of Molecular Sciences. 2024; 25(19):10532. https://doi.org/10.3390/ijms251910532
Chicago/Turabian StyleZhu, Ying, Xinsheng Yang, Yingming Yang, Xu Yan, Chao Li, and Songlin Chen. 2024. "Identification and Functional Analysis of Ras-Related Associated with Diabetes Gene (rrad) in Edwardsiella piscicida-Resistant Individuals of Japanese Flounder (Paralichthys olivaceus)" International Journal of Molecular Sciences 25, no. 19: 10532. https://doi.org/10.3390/ijms251910532