Identification of Insulin-like Growth Factor (IGF) Family Genes in the Golden Pompano, Trachinotus ovatus: Molecular Cloning, Characterization and Gene Expression
Abstract
:1. Introduction
2. Results
2.1. Molecular Cloning and Sequence Analysis
2.2. Tissue Distribution
2.3. Effects of Fasting on IGF1 and IGF2 Expression
2.4. In Vitro Effects of E2 on IGFs mRNA Expression in the Liver and Gonads of Golden Pompano
2.5. In Vitro Effects of MT on IGF mRNA Expression in the Liver and Gonads of Golden Pompano
3. Discussion
4. Materials and Methods
4.1. Experimental Fish Collection
4.2. RNA Extraction and cDNA Synthesis
4.3. Molecular Cloning and Sequence Analysis
4.4. Tissue Distribution of IGF Family Genes
4.5. Effects of Fasting on IGF1 and IGF2 Expression in the Liver and Muscles
4.6. In Vitro Incubation of Golden Pompano with E2 and MT on IGF Gene Expression
4.7. Quantitative Real-Time PCR (RT-qPCR) Analysis
4.8. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | |
---|---|---|---|---|---|---|---|---|---|---|
1: Trachinotus ovatus | 100 | |||||||||
2: Scatophagus argus | 96.76 | 100 | ||||||||
3: Epinephelus coioides | 98.92 | 97.85 | 100 | |||||||
4: Fundulus heteroclitus | 80.87 | 80.98 | 80.98 | 100 | ||||||
5: Larimichthys crocea | 98.92 | 97.85 | 100 | 80.98 | 100 | |||||
6: Nothobranchius furzeri | 88.59 | 87.57 | 88.65 | 82.89 | 88.65 | 100 | ||||
7: Oncorhynchus mykiss | 89.02 | 87.36 | 89.08 | 73.99 | 89.08 | 82.66 | 100 | |||
8: Oreaochromis niloticus | 88.95 | 89.56 | 90.11 | 79.44 | 90.11 | 87.29 | 85.38 | 100 | ||
9: Oryzias latipes | 86.89 | 84.78 | 86.96 | 76.50 | 86.96 | 84.25 | 80.81 | 82.22 | 100 | |
10: Danio rerio | 80.38 | 77.99 | 79.87 | 68.79 | 79.87 | 77.22 | 86.67 | 78.48 | 78.34 | 100 |
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
---|---|---|---|---|---|---|---|---|---|---|---|---|
1: Trachinotus ovatus | 100 | |||||||||||
2: Scatophagus argus | 96.28 | 100 | ||||||||||
3: Epinephelus coioides | 94.88 | 95.81 | 100 | |||||||||
4: Larimichthys crocea | 95.35 | 97.21 | 95.35 | 100 | ||||||||
5: Nothobranchius furzeri | 84.04 | 85.45 | 84.98 | 84.51 | 100 | |||||||
6: Oncorhynchus mykiss | 84.11 | 84.58 | 83.64 | 85.05 | 78.30 | 100 | ||||||
7: Oreaochromis niloticus | 89.77 | 90.70 | 89.30 | 90.23 | 82.16 | 80.84 | 100 | |||||
8: Oryzias latipes | 75 | 74.53 | 75 | 74.06 | 68.10 | 68.72 | 69.81 | 100 | ||||
9: Cyprinus carpio (a) | 52.71 | 53.20 | 53.69 | 53.69 | 52.22 | 54.68 | 53.69 | 46 | 100 | |||
10: Cyprinus carpio (b) | 77.36 | 78.77 | 78.77 | 79.72 | 75.47 | 75.94 | 74.06 | 63.16 | 54.68 | 100 | ||
11: Danio rerio (a) | 55.84 | 56.35 | 56.85 | 57.36 | 55.84 | 58.88 | 56.35 | 47.42 | 83.25 | 55.84 | 100 | |
12: Danio rerio (b) | 77.36 | 78.77 | 78.77 | 79.72 | 75.47 | 76.89 | 74.06 | 63.16 | 54.19 | 95.28 | 55.33 | 100 |
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | |
---|---|---|---|---|---|---|---|---|---|
1: Trachinotus ovatus | 100 | ||||||||
2: Scatophagus argus | 77.60 | 100 | |||||||
3: Epinephelus coioides | 72.02 | 74.62 | 100 | ||||||
4: Fundulus heteroclitus | 59.78 | 62.77 | 59.47 | 100 | |||||
5: Morone saxatilis | 77.20 | 80.20 | 73.87 | 64.21 | 100 | ||||
6: Nothobranchius furzeri | 64.77 | 67.78 | 63.74 | 69.78 | 66.48 | 100 | |||
7: Oreaochromis niloticus | 59.09 | 60.00 | 59.89 | 54.24 | 61.54 | 57.31 | 100 | ||
8: Oryzias latipes | 56.88 | 52.96 | 51.52 | 51.22 | 52.73 | 53.75 | 47.83 | 100 | |
9: Danio rerio | 38.37 | 38.07 | 39.89 | 36.52 | 37.64 | 37.64 | 33.73 | 38.61 | 100 |
Primer Name | Sequence (5′-3′) | Application | Reference |
---|---|---|---|
IGF1 F | GACCCGTGGGGATGTCTAGC | Cloning of ORF | Designed by author |
IGF1 R | ACAGAATGTAGGGAAGGAGC | Cloning of ORF | Designed by author |
IGF2 F | GACTACTGCCATCTGACATG | Cloning of ORF | Designed by author |
IGF2 R | GCACAGACAAGAGTTTGGAG | Cloning of ORF | Designed by author |
IGF3 F | GATGCACTCCTCCTGCCGCTC | Cloning of ORF | Designed by author |
IGF3 R | CGTGACCTCTGACCTCATGGCC | Cloning of ORF | Designed by author |
IGF1 F | TACTGCTGTGTGTCCTCAC | qPCR | Designed by author |
IGF1 R | CGCCTGGAGATGTACTGTGCA | qPCR | Designed by author |
IGF2 F | TCGGCGGAGACGCTGTGTG | qPCR | Designed by author |
IGF2 R | TTCCCGTGATGCCCGCACTA | qPCR | Designed by author |
IGF3 F | GGGGATCGAGGAATCTATT | qPCR | Designed by author |
IGF3 R | AGTTCCAAACAGTATTTCACAA | qPCR | Designed by author |
β-actin F | GAGAGGTTCCGTTGCCCAGAG | qPCR | [2] |
β-actin R | CAGACAGCACAGTGTTGGCGT | qPCR | [2] |
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Ndandala, C.B.; Zhou, Q.; Li, Z.; Guo, Y.; Li, G.; Chen, H. Identification of Insulin-like Growth Factor (IGF) Family Genes in the Golden Pompano, Trachinotus ovatus: Molecular Cloning, Characterization and Gene Expression. Int. J. Mol. Sci. 2024, 25, 2499. https://doi.org/10.3390/ijms25052499
Ndandala CB, Zhou Q, Li Z, Guo Y, Li G, Chen H. Identification of Insulin-like Growth Factor (IGF) Family Genes in the Golden Pompano, Trachinotus ovatus: Molecular Cloning, Characterization and Gene Expression. International Journal of Molecular Sciences. 2024; 25(5):2499. https://doi.org/10.3390/ijms25052499
Chicago/Turabian StyleNdandala, Charles Brighton, Qi Zhou, Zhiyuan Li, Yuwen Guo, Guangli Li, and Huapu Chen. 2024. "Identification of Insulin-like Growth Factor (IGF) Family Genes in the Golden Pompano, Trachinotus ovatus: Molecular Cloning, Characterization and Gene Expression" International Journal of Molecular Sciences 25, no. 5: 2499. https://doi.org/10.3390/ijms25052499