Identification of the Extracellular Nuclease Influencing Soaking RNA Interference Efficiency in Bursaphelenchus xylophilus
Abstract
:1. Introduction
2. Results
2.1. In Vitro Degradation of dsRNA by Nematodes
2.2. Screening of Extracellular Nuclease Genes
2.3. Extracellular Nuclease Gene Response to dsGFP
2.4. The Silencing of Extracellular Nuclease Genes Improves RNAi Efficiency
3. Discussion
4. Materials and Methods
4.1. Cultivation of Nematodes
4.2. Synthesis of dsRNA
4.3. Degradation of dsRNA Detected by Agarose Gel Electrophoresis
4.4. Screening and Cloning of B. xylophilus Extracellular Nuclease Genes
4.5. Interference of Target Gene and Extracellular Nuclease Genes
4.6. Quantitative Reverse Transcription PCR (qRT-PCR)
4.7. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Name | Accession Number | ORF Length | Amino Acid Sequence Length | Signal Peptide | Theoretical pI/Mw |
---|---|---|---|---|---|
BxyNuc1 | OP482151 | 993 | 330 | 20–21.VAS-KS | 5.38/37,069.64 |
BxyNuc2 | OP482152 | 1068 | 355 | 21–22.AEA-AG | 6.37/40,630.98 |
BxyNuc3 | OP482153 | 1035 | 344 | 20–21.VAA-SH | 7.26/39,173.47 |
BxyNuc4 | OP482154 | 933 | 310 | 17–18.GNA-AI | 8.38/34,944.08 |
BxyNuc5 | OP482155 | 890 | 296 | 15–16.TSA-QI | 8.85/33,692.55 |
BxyNuc6 | OP482156 | 927 | 308 | 18–19.ICG-TQ | 9.12/35,178.94 |
BxyNuc7 | OP482157 | 963 | 320 | 19–20.AQG-IG | 5.72/36,009.80 |
BxyNuc8 | OP482158 | 933 | 310 | 17–18.GNA-VI | 8.58/34,493.57 |
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Wang, R.; Li, Y.; Li, D.; Zhang, W.; Wang, X.; Wen, X.; Liu, Z.; Feng, Y.; Zhang, X. Identification of the Extracellular Nuclease Influencing Soaking RNA Interference Efficiency in Bursaphelenchus xylophilus. Int. J. Mol. Sci. 2022, 23, 12278. https://doi.org/10.3390/ijms232012278
Wang R, Li Y, Li D, Zhang W, Wang X, Wen X, Liu Z, Feng Y, Zhang X. Identification of the Extracellular Nuclease Influencing Soaking RNA Interference Efficiency in Bursaphelenchus xylophilus. International Journal of Molecular Sciences. 2022; 23(20):12278. https://doi.org/10.3390/ijms232012278
Chicago/Turabian StyleWang, Ruijiong, Yongxia Li, Dongzhen Li, Wei Zhang, Xuan Wang, Xiaojian Wen, Zhenkai Liu, Yuqian Feng, and Xingyao Zhang. 2022. "Identification of the Extracellular Nuclease Influencing Soaking RNA Interference Efficiency in Bursaphelenchus xylophilus" International Journal of Molecular Sciences 23, no. 20: 12278. https://doi.org/10.3390/ijms232012278