ZFIN ID: ZDB-PUB-091215-15
Transgenic zebrafish eggs containing bactericidal peptide is a novel food supplement enhancing resistance to pathogenic infection of fish
Lin, C.Y., Yang, P.H., Kao, C.L., Huang, H.I., and Tsai, H.J.
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.
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.
ADDITIONAL INFORMATION