Unlocking the Transcriptional Control of NCAPG in Bovine Myoblasts: CREB1 and MYOD1 as Key Players
Abstract
:1. Introduction
2. Results
2.1. The Isolation and Culture of Bovine Myoblasts
2.2. Identification of Bovine Myoblasts
2.3. Structure Analysis of Bovine NCAPG and Phylogenetic Tree Construction
2.4. Schema of NCAPG Expression in Bovine Tissues and Organs
2.5. Confirm of Bovine NCAPG Core Promoter Region
2.6. Validation of the TF Functions
2.7. DNA–Protein Interactions through Electrophoretic Mobility Shift Assays
2.8. Effect of NCAPG on Genes Involved in Myogenesis in Bovine Myoblasts
3. Discussion
4. Materials and Methods
4.1. Sample Collection and Preparation
4.2. Isolation, Identification, and Culture Bovine Myoblasts
4.3. DNA, RNA Isolation, and mRNA Expression Analysis
4.4. Promoter Cloning and Sequence Structure Analysis
4.5. Construction of Promoter Luciferase Reporter Vectors
4.6. Cell Culture, Transfection, and Relative Activity of Dual-Luciferase Assay
4.7. Site-Directed Mutagenesis
4.8. MYOD1, CREB1, and NCAPG Overexpression
4.9. Electrophoretic Mobility Shift Assays
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Chen, Z.; Li, J.; Bai, Y.; Liu, Z.; Wei, Y.; Guo, D.; Jia, X.; Shi, B.; Zhang, X.; Zhao, Z.; et al. Unlocking the Transcriptional Control of NCAPG in Bovine Myoblasts: CREB1 and MYOD1 as Key Players. Int. J. Mol. Sci. 2024, 25, 2506. https://doi.org/10.3390/ijms25052506
Chen Z, Li J, Bai Y, Liu Z, Wei Y, Guo D, Jia X, Shi B, Zhang X, Zhao Z, et al. Unlocking the Transcriptional Control of NCAPG in Bovine Myoblasts: CREB1 and MYOD1 as Key Players. International Journal of Molecular Sciences. 2024; 25(5):2506. https://doi.org/10.3390/ijms25052506
Chicago/Turabian StyleChen, Zongchang, Jingsheng Li, Yanbin Bai, Zhanxin Liu, Yali Wei, Dashan Guo, Xue Jia, Bingang Shi, Xiaolan Zhang, Zhidong Zhao, and et al. 2024. "Unlocking the Transcriptional Control of NCAPG in Bovine Myoblasts: CREB1 and MYOD1 as Key Players" International Journal of Molecular Sciences 25, no. 5: 2506. https://doi.org/10.3390/ijms25052506