Molecular cloning of an IL-8-like CXC chemokine and tissue factor in rainbow trout (Oncorhynchus mykiss) by use of suppression subtractive hybridization.
Suppression subtractive hybridization (SSH) was performed to construct a Rainbow trout cDNA library enriched in sequences up-regulated in head kidney leukocytes after lipopolysaccharide (LPS) and tumour necrosis factor alpha (TNFalpha) stimulation. Random sequencing of fifty clones allowed the identification of a Rainbow trout interleukin 8 (IL-8)-related CXC chemokine, as well as the Rainbow trout tissue factor (TF) precursor. Expression of both the IL-8-like chemokine and TF is induced after LPS and TNFalpha stimulation, indicating that they are associated with inflammatory responses in fish, as has been suggested in mammals. These results confirm the potential of SSH to identify cytokines and immuno-regulatory genes in fish.
Sangrador-Vegas
Amaia
A
National Diagnostics Centre (BioResearch Ireland), National University of Ireland, University Road, Galway, Ireland.
Lennington
Jessica B
JB
Smith
Terry J
TJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019743:Chemokines, CXC / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D029941:Fish Proteins / Q000235:genetics*; D016209:Interleukin-8 / Q000235:genetics*; D007668:Kidney / Q000187:drug effects / Q000276:immunology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods; D017686:Oncorhynchus mykiss / Q000235:genetics*; D013925:Thromboplastin / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D015854:Up-Regulation
D019743:Chemokines, CXC; D018076:DNA, Complementary; D029941:Fish Proteins; D016209:Interleukin-8; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D013925:Thromboplastin