RNA Interference of Chitin Synthase 2 Gene in Liriomyza trifolii through Immersion in Double-Stranded RNA
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Insects
2.2. Cloning, Sequence Alignment, and Expression of CHS2 Transcript
2.3. Synthesis and Delivery of dsRNA
2.4. Analysis of Silencing Efficiency
2.5. Statistical analysis
3. Results
3.1. Sequence Characteristics and Phylogenetic Analysis of CHS2 Transcript from L. trifolii
3.2. Expression Pattern of CHS2 Transcript in L. trifolii
3.3. Functional Verification of CHS2 Transcript of L. trifolii
4. Discussion
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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Gene | Primer Sequences (5′→3′) | Length (bp) | Tm (°C) | ||
Primers for cDNA cloning and full-length cDNA amplification | |||||
CHS2 | F | TACACACTGAGGGTTCCCATTT | 999 | 53.9 | |
R | GAACTCATTGAAACCTTCTGGA | ||||
5′ | TTGTCATCGCTCTCACCCGCTCC | 393 | 64.5 | ||
3′ | CTGCGAATCTGACAGGGCGTTTG | 1566 | 61.6 | ||
Primers for dsRNA synthesis | |||||
dsCHS2 | F | TAATACGACTCACTATAGGGTTAGTTTGGATTTTACCTGGCA | 553 | 63.1 | |
R | TAATACGACTCACTATAGGGTTCTACACGATAGCCTCTTTGC | ||||
dsGFP | F | TAATACGACTCACTATAGGGCCTCGTGACCACCCTGACCTAC | 314 | 68.2 | |
R | TAATACGACTCACTATAGGGCACCTTGATGCCGTTCTTCTGC | ||||
Primers for qRT-PCR | Length (bp) | Tm (°C) | Efficiency (%) | ||
qPCR-CHS2 | F | TACACACTGAGGGTTCCCATTTA | 116 | 59.0 | 97.5 |
R | TCCATCATTTCATCCTTAGTTTC | ||||
qPCR-Actin | F | TTGTATTGGACTCTGGTGACGG | 73 | 59.2 | 108.6 |
R | GATAGCGTGAGGCAAAGCATAA | ||||
qPCR-18S | F | GAAGCAGTTTGGGGGCATTA | 88 | 60.0 | 103.2 |
R | TTGGCAAATGCTTTCGCTTA |
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Chang, Y.-W.; Wang, Y.-C.; Yan, Y.-Q.; Xie, H.-F.; Yuan, D.-R.; Du, Y.-Z. RNA Interference of Chitin Synthase 2 Gene in Liriomyza trifolii through Immersion in Double-Stranded RNA. Insects 2022, 13, 832. https://doi.org/10.3390/insects13090832
Chang Y-W, Wang Y-C, Yan Y-Q, Xie H-F, Yuan D-R, Du Y-Z. RNA Interference of Chitin Synthase 2 Gene in Liriomyza trifolii through Immersion in Double-Stranded RNA. Insects. 2022; 13(9):832. https://doi.org/10.3390/insects13090832
Chicago/Turabian StyleChang, Ya-Wen, Yu-Cheng Wang, Yu-Qing Yan, Hong-Fang Xie, Deng-Rong Yuan, and Yu-Zhou Du. 2022. "RNA Interference of Chitin Synthase 2 Gene in Liriomyza trifolii through Immersion in Double-Stranded RNA" Insects 13, no. 9: 832. https://doi.org/10.3390/insects13090832