Functional Analysis of a CTL-X-Type Lectin CTL16 in Development and Innate Immunity of Tribolium castaneum
Abstract
:1. Introduction
2. Results
2.1. Domain Organization, Structure, and Evolutionary Relationships among CTLs
2.2. Spatiotemporal Expression of TcCTL16
2.3. TcCTL16 Silencing in Early Larvae Influence Growth and Development
2.4. TcCTL16 Responses to Polysaccharide or Bacterial Challenge
2.5. Production of rTcCTL16 CRD and Its Binding and Agglutination Capacity to Microbes
2.6. TcCTL16 Silencing on Late Larvae Does Not Affect Viability of Beetles
2.7. Effects of TcCTL16 Gene Knockdown on Defense against Bacterial Infection
3. Discussion
4. Materials and Methods
4.1. Animals and Chemicals
4.2. Identification and Cloning of TcCTL16 Gene
4.3. Sequence and Phylogenetic Analysis
4.4. Spatiotemporal Expression of TcCTL16
4.5. Expression Profiles of TcCTL16 after Being Challenged with Polysaccharides and Bacteria
4.6. Production of Recombinant TcCTL16 and Western Blot Analysis
4.7. Microorganism Binding and Agglutination of TcCTL16
4.8. dsRNA-Mediated RNAi Assay
4.9. Survival Assay
4.10. Statistical Analyses
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Name | Sequence (5′-3′) |
---|---|
PCR: | |
TcCTL16-F | ATGGTCGTGTGTCAGTGCAAGG |
TcCTL16-R | ATATCTCTTCTGGTCGCGGTGC |
TcCTL16-F | atgggtcgcggatccgaattcATGGTCGTGTGTCAGTGCAAGG |
TcCTL16-R | gtggtggtggtggtgctcgagATATCTCTTCTGGTCGCGGTGC |
RNAi: | |
ds-TcCTL16-F | taatacgactcactatagggATGGTCGTGTGTCAGTGCAA |
ds-TcCTL16-R | taatacgactcactatagggATTGTGGCAGGTTTGTTCGC |
qRT-PCR: | |
TcCTL16-qF | TGTGAACCTGTCCAGTGCG |
TcCTL16-qR | CATCATACTGTCCATTCGCC |
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Bi, J.; Wang, Y.; Gao, R.; Liu, P.; Jiang, Y.; Gao, L.; Li, B.; Song, Q.; Ning, M. Functional Analysis of a CTL-X-Type Lectin CTL16 in Development and Innate Immunity of Tribolium castaneum. Int. J. Mol. Sci. 2023, 24, 10700. https://doi.org/10.3390/ijms241310700
Bi J, Wang Y, Gao R, Liu P, Jiang Y, Gao L, Li B, Song Q, Ning M. Functional Analysis of a CTL-X-Type Lectin CTL16 in Development and Innate Immunity of Tribolium castaneum. International Journal of Molecular Sciences. 2023; 24(13):10700. https://doi.org/10.3390/ijms241310700
Chicago/Turabian StyleBi, Jingxiu, Yutao Wang, Rui Gao, Pingxiang Liu, Yuying Jiang, Lei Gao, Bin Li, Qisheng Song, and Mingxiao Ning. 2023. "Functional Analysis of a CTL-X-Type Lectin CTL16 in Development and Innate Immunity of Tribolium castaneum" International Journal of Molecular Sciences 24, no. 13: 10700. https://doi.org/10.3390/ijms241310700