PUBLICATION
Transgenic zebrafish eggs containing bactericidal peptide is a novel food supplement enhancing resistance to pathogenic infection of fish
- Authors
- Lin, C.Y., Yang, P.H., Kao, C.L., Huang, H.I., and Tsai, H.J.
- ID
- ZDB-PUB-091215-15
- Date
- 2010
- Source
- Fish & shellfish immunology 28(3): 419-427 (Journal)
- Registered Authors
- Tsai, Huai-Jen
- Keywords
- Bactericide, Embryos, Feeding, Lactoferrincin, Transgenic, Zebrafish
- MeSH Terms
-
- Actins/genetics
- Animals
- Animals, Genetically Modified
- Anti-Bacterial Agents/pharmacology
- Bioreactors
- Blotting, Western
- Cattle
- Dietary Supplements*
- Eggs*
- Fish Diseases/immunology*
- Fish Diseases/mortality
- Fish Diseases/prevention & control
- Gram-Negative Bacteria/drug effects
- Gram-Negative Bacterial Infections/immunology
- Gram-Negative Bacterial Infections/mortality
- Gram-Negative Bacterial Infections/prevention & control
- Gram-Negative Bacterial Infections/veterinary*
- Immunity, Innate/drug effects*
- Lactoferrin*/genetics
- Lactoferrin*/pharmacology
- Polymerase Chain Reaction
- Recombinant Proteins/pharmacology
- Zebrafish*/genetics
- Zebrafish*/metabolism
- Zebrafish*/microbiology
- PubMed
- 19944764 Full text @ Fish Shellfish Immunol.
Citation
Lin, C.Y., Yang, P.H., Kao, C.L., Huang, H.I., and Tsai, H.J. (2010) Transgenic zebrafish eggs containing bactericidal peptide is a novel food supplement enhancing resistance to pathogenic infection of fish. Fish & shellfish immunology. 28(3):419-427.
Abstract
Zebrafish (Danio rerio) was used as a bioreactor to produce bovine lactoferricin (LFB), which has wide-ranging antimicrobial activity. We constructed an expression plasmid in which LFB was fused with green fluorescent protein (GFP) and driven by zebrafish beta-actin promoter. After microinjection, six transgenic founders were screened on the basis of GFP appearance. Among them, a stable ZBL-5 line was selected by the ubiquitous and strong expression of GFP. Using PCR and Western blot analysis, we confirmed that the recombinant LFB-GFP protein was produced by the F2 progeny derived from the ZBL-5 line. The bactericidal agar plate assay proved that the functional domain of LFB was released from the LFB-GFP fusion protein, resulting in strong bactericidal activity against Escherichia coli, Edwardsiella tarda and Aeromonas hydrophila. Furthermore, adult zebrafish were given one feeding of fifty 72-hpf transgenic embryos. The treated fish were then immersed in freshwater containing 1 x 10(5) CFU/ml E. tarda for 7 days. The survival rate of the treated zebrafish was significantly higher than that of fish fed with fifty wild-type embryos (75 +/- 12.5% versus 4 +/- 7.2%). This line of evidence suggested that pathogen resistance can be enhanced by using transgenic embryos containing LFB-GFP as a food supplement for fish, while, at the same time, reducing the demand of chemical antibiotics.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping