Oral Administration of Lactobacillus Casei Expressing Flagellin A Protein Confers Effective Protection against Aeromonas Veronii in Common Carp, Cyprinus Carpio
Abstract
:1. Introduction
2. Results
2.1. Construction of the Recombinant L. casei
2.2. Western Blotting
2.3. Surface Expression of the Modified FlaA
2.4. Humoral Immune Parameters
2.5. Expression of Immune-Related Genes
2.6. Colonization of Recombinant L. casei in the Fish Intestine
2.7. Challenge Test
3. Discussion
4. Materials and Methods
4.1. Fish and Ethics Statement
4.2. Bacterial Strains, Plasmids and Growth Conditions
4.3. Construction of Recombinant L. casei Expressing FlaA Gene
4.4. Western Blotting Assay
4.5. Expression of FlaA on the Cell Surface
4.6. Oral Immunization and Sample Collection
4.7. Phagocytic Activity of Leukocyte
4.8. Enzyme-Linked Immunosorbent Assay (ELISA)
4.9. Real-Time PCR Analysis
4.10. Colonization of Recombinant L. casei in the Fish Intestine
4.11. Challenge Test
4.12. Data Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Primer | Sequence(5’-3’) | Accession | PCR Product (bp) |
---|---|---|---|
IL-8 | F: CATTCAGAGCCAGCAATT | AB470924.1 | 135 |
R: CACCCAGTCTTTAGTAGGAT | |||
IL-10 | F: AACTGATGACCCGAATGGAAAC | JX524550.1 | 143 |
R: CACCTTCTCCCAGTCGTCAAA | |||
IL-1β | F: CATCAAAGAAATCGCTCCTG | AB010701 | 133 |
R: GCAAGGTCTGCCTGGTCT | |||
TNF-α | F: TTATGTCGGTGCGGCCTTC | AJ311800.2 | 101 |
R: AGGTCTTTCCGTTGTCGCTTT | |||
IFN-γ | F: AACAGTCGGGTGTCGCAAG | AB376666.1 | 141 |
R: TCAGCAAACATACTCCCCAG | |||
β-actin | F: CAAGATGATGGTGTGCCAAGTG | M24113.1 | 352 |
R: TCTGTCTCCGGCACGAAGTA | |||
pPG-1-FlaA | F:AACCCGGGATGGGCCTTTTTATCAACACTAACG | CP012504.1 | 903 |
R:CCGATATCTTAGCCTTGCAGCAGCTGAAGTGCT | |||
pPG-2-FlaA | F: CGGGATCCATGGGCCTTTTTATCAACACTAACG | CP012504.1 | 903 |
R:CCCTCGAGTTAGCCTTGCAGCAGCTGAAGTGCT |
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Tian, J.-X.; Kang, Y.-H.; Chu, G.-S.; Liu, H.-J.; Kong, Y.-D.; Zhao, L.-H.; Kong, Y.-X.; Shan, X.-F.; Wang, G.-Q. Oral Administration of Lactobacillus Casei Expressing Flagellin A Protein Confers Effective Protection against Aeromonas Veronii in Common Carp, Cyprinus Carpio. Int. J. Mol. Sci. 2020, 21, 33. https://doi.org/10.3390/ijms21010033
Tian J-X, Kang Y-H, Chu G-S, Liu H-J, Kong Y-D, Zhao L-H, Kong Y-X, Shan X-F, Wang G-Q. Oral Administration of Lactobacillus Casei Expressing Flagellin A Protein Confers Effective Protection against Aeromonas Veronii in Common Carp, Cyprinus Carpio. International Journal of Molecular Sciences. 2020; 21(1):33. https://doi.org/10.3390/ijms21010033
Chicago/Turabian StyleTian, Jia-Xin, Yuan-Huan Kang, Guo-Sheng Chu, Hong-Jian Liu, Yi-Di Kong, Lin-Hui Zhao, Yu-Xin Kong, Xiao-Feng Shan, and Gui-Qin Wang. 2020. "Oral Administration of Lactobacillus Casei Expressing Flagellin A Protein Confers Effective Protection against Aeromonas Veronii in Common Carp, Cyprinus Carpio" International Journal of Molecular Sciences 21, no. 1: 33. https://doi.org/10.3390/ijms21010033