Molecular Characterization and Virus-Induced Gene Silencing of a Collagen Gene, Me-col-1, in Root-Knot Nematode Meloidogyne enterolobii
Abstract
:1. Introduction
2. Materials and Methods
2.1. Nematodes and Plants
2.2. Cloning of Me-col-1 cDNA Sequence
2.3. Bioinformatic Analysis of Me-col-1 Sequence
2.4. Developmental Expression Analysis
2.5. Heterologous Expression of Protein Me-col-1
2.6. dsRNA Preparation and In Vitro RNAi of Me-col-1
2.7. In Planta RNAi
2.8. Statistical Analysis
3. Results
3.1. Sequence Characterization and Analysis of Me-col-1 Gene
3.2. Prokaryotic Expression of Me-col-1 Protein
3.3. Temporal Expression Assays
3.4. Me-col-1 In Vitro RNAi
3.5. Me-col-1 In Vivo RNAi
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Primer Name | Primer Sequences (5′–3′) |
---|---|
GeneRacer5′ primer | CGACTGGAGCACGAGGACACTGA |
C-S1 | CCCTGTCCATAGGTTGCCCCAC |
GeneRacer5′ nested primer | GGACACTGACATGGACTGAAGGAGTA |
C-S2 | TCCAGGCACGGGAATGAGACGA |
GeneRacer3′ primer | TACCGTCGTTCCACTAGTGATTT |
M-A1 | GGGGCAACCTATGGACAGGGAGC |
GeneRacer3′ nested primer | CGCGGATCCTCCACTAGTGATTTCACTATAGG |
M-A2 | CTGGTCGTCTCATTCCCGTGCCT |
Col-F | ATGGAACCTAAAGAGCAG |
Col-R | TTAATGATATCCACCACTTTTTGCACTTGC |
Col-QF | GCTCTCCTGGTCGTCTCATT |
Col-QR | ACAACTGCCCTTATCTCCG |
MeActF | ACGGTCAAGTCATTACTGTTGGAAA |
MeActR | GTAAAGGTCTTTACGGATGTCAATG |
Col-BamHI | CGGATCCGAACCTAAAGAGCAGTTTTGC |
Col-NotI | TTGCGGCCGCAATGATATCCACCACTTTTTGCACT |
Col-IF 1 | TAATACGACTCACTATAGGGAGATGTAGCGGTTGTTGTTTCG |
Col-IR 1 | TAATACGACTCACTATAGGGAGATTTCCATTAGGTCCATCCTC |
TRVF | CTGGGTTACTAGCGGCACTGAATA |
TRVR | TCCACCAAACTTAATCCCGAATAC |
DcolF | TGTAGCGGTTGTTGTTTCG |
DcolR | TTTCCATTAGGTCCATCCTC |
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Pei, J.; Feng, T.; Long, H.; Chen, Y.; Pei, Y.; Sun, Y. Molecular Characterization and Virus-Induced Gene Silencing of a Collagen Gene, Me-col-1, in Root-Knot Nematode Meloidogyne enterolobii. Life 2022, 12, 2103. https://0-doi-org.brum.beds.ac.uk/10.3390/life12122103
Pei J, Feng T, Long H, Chen Y, Pei Y, Sun Y. Molecular Characterization and Virus-Induced Gene Silencing of a Collagen Gene, Me-col-1, in Root-Knot Nematode Meloidogyne enterolobii. Life. 2022; 12(12):2103. https://0-doi-org.brum.beds.ac.uk/10.3390/life12122103
Chicago/Turabian StylePei, Ji, Tuizi Feng, Haibo Long, Yuan Chen, Yueling Pei, and Yanfang Sun. 2022. "Molecular Characterization and Virus-Induced Gene Silencing of a Collagen Gene, Me-col-1, in Root-Knot Nematode Meloidogyne enterolobii" Life 12, no. 12: 2103. https://0-doi-org.brum.beds.ac.uk/10.3390/life12122103