Cloning, Characterization and Effect of TmPGRP-LE Gene Silencing on Survival of Tenebrio Molitor against Listeria monocytogenes Infection
Abstract
:1. Introduction
2. Results and Discussion
2.1. Characterization of TmPGRP-LE Full-Length cDNA
2.2. Phylogenetic Analysis of TmPGRP-LE
2.3. Secondary Structure and Homology Modeling of TmPGRP-LE
2.4. Temporal Expression Patterns of TmPGRP-LE
2.5. RNAi-Mediated Knockdown of TmPGRP-LE Leads to Reduced Survival Rate of T. molitor Larvae infected with L. monocytogenes
3. Experimental Section
3.1. Insect Collection and Maintenance
3.2. Chemicals
3.3. Bacterial Strains and Media
3.4. Construction of Full-Length Enriched cDNA Library of T. molitor Larvae
3.5. TmPGRP-LE Sequence Analysis
3.6. Phylogenetic Analysis
3.7. Secondary Structure Prediction and Modeling
3.8. Developmental Expression Pattern of TmPGRP-LE
3.9. RNA Interference of TmPGRP-LE
3.10. Bacterial Injections and Bioassays
3.11. Statistical Analysis
4. Conclusions
Supplementary Information
ijms-14-22462-s001.pdfPrimer name | Sequence (5′-3′) |
---|---|
Oligo (dT) adaptor | GGCCACGCGTCGACTAGTACT17 |
M13-F (forward) | GTAAAACGACGGCCAG |
M13-R (reverse) | CAGGAAACAGCTATGAC |
RpL27a (forward) | TCATCCTGAAGGCAAAGCTCCAGT |
RpL27a (reverse) | AGGTTGGTTAGGCAGGCACCTTTA |
TmPGRP-LE Real-time PCR (Forward) | CTTCGCTTGCGGAATGGCAGATTA |
TmPGRP-LE Real-time PCR (Reverse) | AACACACGCTCAAATCCTTTCCCG |
TmPGRP-LE dsRNA (forward) | TAATACGACTCACTATAGGGAGAGGCAACGTAAATAAGGACGG |
TmPGRP-LE dsRNA (Reverse) | TAATACGACTCACTATAGGGAGAGTAGGCGATATCGTTCCACTTC |
Acknowledgments
Conflicts of Interest
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Tindwa, H.; Patnaik, B.B.; Kim, D.H.; Mun, S.; Jo, Y.H.; Lee, B.L.; Lee, Y.S.; Kim, N.J.; Han, Y.S. Cloning, Characterization and Effect of TmPGRP-LE Gene Silencing on Survival of Tenebrio Molitor against Listeria monocytogenes Infection. Int. J. Mol. Sci. 2013, 14, 22462-22482. https://doi.org/10.3390/ijms141122462
Tindwa H, Patnaik BB, Kim DH, Mun S, Jo YH, Lee BL, Lee YS, Kim NJ, Han YS. Cloning, Characterization and Effect of TmPGRP-LE Gene Silencing on Survival of Tenebrio Molitor against Listeria monocytogenes Infection. International Journal of Molecular Sciences. 2013; 14(11):22462-22482. https://doi.org/10.3390/ijms141122462
Chicago/Turabian StyleTindwa, Hamisi, Bharat Bhusan Patnaik, Dong Hyun Kim, Seulgi Mun, Yong Hun Jo, Bok Luel Lee, Yong Seok Lee, Nam Jung Kim, and Yeon Soo Han. 2013. "Cloning, Characterization and Effect of TmPGRP-LE Gene Silencing on Survival of Tenebrio Molitor against Listeria monocytogenes Infection" International Journal of Molecular Sciences 14, no. 11: 22462-22482. https://doi.org/10.3390/ijms141122462