Bombyx mori Ecdysone Receptor B1 May Inhibit BmNPV Infection by Triggering Apoptosis
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Silkworms and BmN Cells
2.2. Preparation and Detection of BmNPV
2.3. Sample Preparation for Expression Pattern Analysis
2.4. Bioinformatics Analysis
2.5. RNA Extraction and the First Strand cDNA Synthesis
2.6. Real-Time Quantitative PCR (RT-qPCR)
2.7. Synthesis of siRNA and Transfection
2.8. Construction of the Overexpression Vector
2.9. The Treatment of 20E on Silkworm Larvae
2.10. The Effect of 20E Treatment on BmEcR-B1 Expression and BmNPV Infection in BmN Cells
2.11. Caspase Activity Assay and Apoptotic Bodies Detection
2.12. Statistics Analysis
3. Results
3.1. Bioinformatics Analysis
3.2. The Temporal and Spatial Expression Profile of BmEcR-B1 and BmEcR-A
3.3. BmEcR-B1 Showed a Significant Response to BmNPV Infection
3.4. E activates BmEcR-B1 Expression and Inhibits BmNPV Infection In Vivo
3.5. BmEcR-B1 Plays a Vital Role in 20E Inhibition of BmNPV In Vitro
3.6. Overexpression of BmEcR-B1 Inhibits BmNPV Infection in BmN Cells
3.7. Inhibition of Viral Infection by BmEcR-B1 Is Associated with the Activation of Apoptosis
3.8. The Analysis of the Effect of 20E on Silkworm Larvae Development and Cocoon Shell
4. Discussion
4.1. BmEcR-B1 Is More Critical Than BmEcR-A in Silkworm
4.2. BmEcR-B1 Plays a Vital Role in Inhibiting BmNPV, Which Needs the Presence of 20E
4.3. BmEcR-B1 Likely Inhibits Virus Infection in BmN Cells via the Activation of Apoptosis
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Gene Name | Forward Primer (5′-3′) | Reverse Primer (5′-3′) | Note |
---|---|---|---|
BmEcR-B1-O | CGGAATTCATGGACAGAGCCTCCGGATAC | GCTCTAGAGATGCACATGTTGGAGTTTTG | ORF amplification |
BmEcR-B1 | GCCTCCCACAACACCGAAATCA | TCTGGCGTCAGCATCAGCACT | 98.30%, 106 bp |
BmEcR-A | ACTCGTCGCCACTATCCTCAGG | TGAAGCCGGCGAAAGTTCCT | 98.48%, 110 bp |
BmGAPDH | CCGCGTCCCTGTTGCTAAT | CTGCCTCCTTGACCTTTTGC | 116.00%, 98 bp |
vp39 | CAACTTTTTGCGAAACGACTT | GGCTACACCTCCACTTGCTT | 100.71%, 125 bp |
Primer Names | Sequences (5′-3′) |
---|---|
EcR-Olig1-1 | GATCACTAATACGACTCACTATAGGGAATTTGGACATGCCTGTGAAATT |
EcR-Olig1-2 | AATTTCACAGGCATGTCCAAATTCCCTATAGTGAGTCGTATTAGTGATC |
EcR-Olig1-3 | AAAATTTGGACATGCCTGTGAAACCCTATAGTGAGTCGTATTAGTGATC |
EcR-Olig1-4 | GATCACTAATACGACTCACTATAGGGTTTCACAGGCATGTCCAAATTTT |
EcR-Olig2-1 | GATCACTAATACGACTCACTATAGGGAACACGTTGCGAATTTACATCTT |
EcR-Olig2-2 | AAGATGTAAATTCGCAACGTGTTCCCTATAGTGAGTCGTATTAGTGATC |
EcR-Olig2-3 | AAAACACGTTGCGAATTTACATCCCCTATAGTGAGTCGTATTAGTGATC |
EcR-Olig2-4 | GATCACTAATACGACTCACTATAGGGGATGTAAATTCGCAACGTGTTTT |
RFP-Olig-1 | GATCACTAATACGACTCACTATAGGGGCACCCAGACCATGAGAATTT |
RFP-Olig-2 | AAATTCTCATGGTCTGGGTGCCCCTATAGTGAGTCGTATTAGTGATC |
RFP-Olig-3 | AAGCACCCAGACCATGAGAATCCCTATAGTGAGTCGTATTAGTGATC |
RFP-Olig-4 | GATCACTAATACGACTCACTATAGGGATTCTCATGGTCTGGGTGCTT |
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Su, Z.; Zhao, C.; Huang, X.; Lv, J.; Zhao, Z.; Zheng, K.; Sun, X.; Qin, S.; Wang, X.; Jin, B.-R.; et al. Bombyx mori Ecdysone Receptor B1 May Inhibit BmNPV Infection by Triggering Apoptosis. Insects 2023, 14, 505. https://doi.org/10.3390/insects14060505
Su Z, Zhao C, Huang X, Lv J, Zhao Z, Zheng K, Sun X, Qin S, Wang X, Jin B-R, et al. Bombyx mori Ecdysone Receptor B1 May Inhibit BmNPV Infection by Triggering Apoptosis. Insects. 2023; 14(6):505. https://doi.org/10.3390/insects14060505
Chicago/Turabian StyleSu, Zhihao, Chunxiao Zhao, Xinming Huang, Junli Lv, Ziqin Zhao, Kaiyi Zheng, Xia Sun, Sheng Qin, Xueyang Wang, Byung-Rae Jin, and et al. 2023. "Bombyx mori Ecdysone Receptor B1 May Inhibit BmNPV Infection by Triggering Apoptosis" Insects 14, no. 6: 505. https://doi.org/10.3390/insects14060505