Identification and Characterization of a Fatty Acid- and Retinoid-Binding Protein Gene (Ar-far-1) from the Chrysanthemum Foliar Nematode, Aphelenchoides ritzemabosi
Abstract
:1. Introduction
2. Results
2.1. Cloning and Analysis of Full-Length Ar-far-1 from CFN
2.2. Southern Blot Analysis
2.3. Ligand Binding
2.4. Localization and Expression of Ar-far-1 mRNA
2.5. Ar-far-1 Silencing Efficiency and Influence on the Reproduction and Pathogenicity of CFN
3. Discussion
4. Materials and Methods
4.1. Nematodes
4.2. RNA Extraction and Cloning of Ar-far-1 from CFN
4.3. Sequence Analysis, Alignment and Phylogenetics
4.4. Southern Blot Hybridization
4.5. Expression and Purification of Recombinant Ar-FAR-1 and Fluorescence-Based Ligand-Binding Assays
4.6. In Situ Hybridization
4.7. Expression of Ar-far-1 mRNA at Different Developmental Stages of CFN
4.8. Reproduction and Pathogenicity of CFN after RNAi Treatment
4.9. Data Analysis
Data Availability Statement
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Primer Name | Sequence | Primer Use |
---|---|---|
FAR1-F | 5′-AACCCAAGTTTGAGCAACCTC-3′ | cDNA and gDNA amplification |
FAR1-R | 5′-TATGAAGTTTATTTCGTGTTT-3′ | |
FAR1-Sb-F | 5′-AGAAGTTGTGCCCGAGGAAGT-3′ | Southern blot |
FAR1-Sb-R | 5′-ACCTTCAACAAATGCTTTCGC-3′ | |
FAR1FBamHI | 5′-CGGGATCCATGAGTCTTCGCACATTCGTT-3′ | Prokaryotic expression |
FAR1RXhoI | 5′-CCGCTCGAGTTAATTTTCTTGTTTGATCAA-3′ | |
FAR1-IN-T7F | 5′-GGATCCTAATACGACTCACTATAGGGTACTCTTCCGTTGAGCATTC-3′ | In situ hybridization and dsRNA template |
FAR1-IN-R | 5′- GCTTCTCTTCACCCTTAGGT-3′ | |
FAR1-IN-F | 5′- TACTCTTCCGTTGAGCATTC-3′ | |
FAR1-IN-T7R | 5′-GGATCCTAATACGACTCACTATAGGGGCTTCTCTTCACCCTTAGGT-3′ | |
qFAR1-F | 5′-GTCTTCGCACATTCGTTTC-3′ | qRT-PCR |
qFAR1-R | 5′-ATTTTTTCACTTCCTCGGG-3′ | |
Y18s-F | 5′-GACTCAACACGGGAAACCTCA-3′ | qRT-PCR |
Y18s-R | 5′-GCAGACACTCCACACAAGCAC-3′ |
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Ding, S.-W.; Wang, D.-W.; Xiang, Y.; Xu, C.-L.; Xie, H. Identification and Characterization of a Fatty Acid- and Retinoid-Binding Protein Gene (Ar-far-1) from the Chrysanthemum Foliar Nematode, Aphelenchoides ritzemabosi. Int. J. Mol. Sci. 2019, 20, 5566. https://doi.org/10.3390/ijms20225566
Ding S-W, Wang D-W, Xiang Y, Xu C-L, Xie H. Identification and Characterization of a Fatty Acid- and Retinoid-Binding Protein Gene (Ar-far-1) from the Chrysanthemum Foliar Nematode, Aphelenchoides ritzemabosi. International Journal of Molecular Sciences. 2019; 20(22):5566. https://doi.org/10.3390/ijms20225566
Chicago/Turabian StyleDing, Shan-Wen, Dong-Wei Wang, Yu Xiang, Chun-Ling Xu, and Hui Xie. 2019. "Identification and Characterization of a Fatty Acid- and Retinoid-Binding Protein Gene (Ar-far-1) from the Chrysanthemum Foliar Nematode, Aphelenchoides ritzemabosi" International Journal of Molecular Sciences 20, no. 22: 5566. https://doi.org/10.3390/ijms20225566