Recombinant Mosquito Densovirus with Bti Toxins Significantly Improves Pathogenicity against Aedes albopictus
Abstract
:1. Introduction
2. Results
2.1. Expression, Cloning, and Identification of Bti Toxins in AeDNV
2.2. Efficiency of Recombinant Virus in C6/36 Cell Lines and Insecticidal Efficacy against Ae. albopictus
2.3. Stability of the Recombinant Viruses
3. Discussion
4. Materials and Methods
4.1. Mosquito Maintenance
4.2. Construction of Recombinant Plasmids
4.3. Cell line Transfection
4.4. Production of Recombinant Virus
4.5. Western Blotting and Nano LC-MS/MS Analysis
4.6. Toxicity Bioassay
4.7. Determination of Genetic Stability of the Virus
4.8. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Sample | LC50 (Copies/mL) | LT50 (Days) | ||||
---|---|---|---|---|---|---|
Copies/mL | 95% CI | 1 × 1010 Copies/mL, 95% CI | 1 × 1011 Copies/mL, 95% CI | |||
AeDNV | 1010.6 | 1010.09–1011.2 | 8.7 d | (8.14–9.08) | 7.28 d | (6.55–7.98) |
α8-AeDNV | 109.8 | 109.39–1010.5 | 7.61 d | (7.26–7.94) | 6.49 d | (5.48–7.48) |
α8Cyt-AeDNV | 109.3 | 108.9–109.93 | 6.71 d | (5.48–7.87) | 5.85 d | (4.61–6.96) |
Days | wt-AeDNV | α8-AeDNV | α8Cyt-AeDNV |
---|---|---|---|
1 | 1 × 108 | 1 × 108 | 1 × 108 |
2 | 2.93 × 108 | 3.21 × 108 | 2.86 × 108 |
3 | 3.12 × 108 | 4.63 × 108 | 3.1 × 108 |
4 | 5.75 × 108 | 5.56 × 108 | 8.55 × 108 |
5 | 8.15 × 108 | 8.24 × 108 | 1.21 × 109 |
6 | 1.56 × 109 | 1.18 × 109 | 1.76 × 109 |
7 | 7.73 × 109 | 4.19 × 109 | 1.1 × 1010 |
9 | 9.24 × 109 | 7.93 × 109 | 8.42 × 1010 |
11 | 1.49 × 1012 | 9.14 × 1011 | 2.41 × 1012 |
Primer Name | Sequences (5′ to 3′) | Annealing Temperature and Cycles | Product Size (bp) | Program Application |
---|---|---|---|---|
Loop α8 F | TTAGCCCCACTCATTGTAGTTGAC | 55 °C, 35 | 255 bp | RT-PCR |
Loop α8 R | CGAATGAGTACAAAACTACACCTC | |||
Loop α8Cyt F | TTGACGGCGGATACTTTGAT | 275 bp | ||
Loop α8Cyt R | CGAATGAGTACAAAACTACACCTC | |||
β-Actin F | CACCAGGGTGTGATGGTCGG | 911 bp | ||
β-Actin R | CCACCGATCCAGACGGAGT |
Primer Name | Sequences (5′ to 3′) | Annealing Temperature and Cycles | Program Application |
---|---|---|---|
AeDNV-qF | AACCGATAGAACGAACAC | 55 °C, 40 | Virus quantification |
AeDNV-qR | TTGGAGGACGACTGATTA | ||
pMDV-F | AACTACCAGGAGCAGGAT | 55 °C, 30 | PCR-based viral genome detection |
pMDV-R | TGTATGTGCGTTGTCTTC |
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Batool, K.; Alam, I.; Liu, P.; Shu, Z.; Zhao, S.; Yang, W.; Jie, X.; Gu, J.; Chen, X.-G. Recombinant Mosquito Densovirus with Bti Toxins Significantly Improves Pathogenicity against Aedes albopictus. Toxins 2022, 14, 147. https://doi.org/10.3390/toxins14020147
Batool K, Alam I, Liu P, Shu Z, Zhao S, Yang W, Jie X, Gu J, Chen X-G. Recombinant Mosquito Densovirus with Bti Toxins Significantly Improves Pathogenicity against Aedes albopictus. Toxins. 2022; 14(2):147. https://doi.org/10.3390/toxins14020147
Chicago/Turabian StyleBatool, Khadija, Intikhab Alam, Peiwen Liu, Zeng Shu, Siyu Zhao, Wenqiang Yang, Xiao Jie, Jinbao Gu, and Xiao-Guang Chen. 2022. "Recombinant Mosquito Densovirus with Bti Toxins Significantly Improves Pathogenicity against Aedes albopictus" Toxins 14, no. 2: 147. https://doi.org/10.3390/toxins14020147