Molecular cloning and expression of canine interleukin 8 cDNA.
Molecular cloning of canine interleukin-8 (IL-8) was performed to establish a basis for its investigation in the canine immune system. From a cDNA pool constructed from LPS-stimulated popliteal lymph node cells, canine IL-8 cDNA covering the whole coding region was amplified by polymerase chain reaction. The nucleotide sequence of a canine IL-8 clone, designated pcIL-8#38, was highly similar to those of human, rabbit and porcine IL-8, and comprised 353 bp with an open reading frame that encoded 101 amino acids. Analysis of the deduced amino acid sequence of insert DNA in pcIL-8#38 showed 76.5, 80.2, and 87.0% similarities with human, rabbit and porcine IL-8 proteins, respectively. Insert DNA of pcIL-8#38 was transferred to a mammalian expression vector, pcDL-SR alpha 296, and transfected into Cos7 cells. The supernatant of the transfectant had neutrophil chemotactic activity when it was examined by the neutrophil migration assay, suggesting that our cloned cDNA was biologically active. The cloned canine IL-8 cDNA will be useful for canine inflammatory disease and comparative immunology research.
Matsumoto
Y
Y
Laboratory of Immune Cytology, National Institute of Animal Health, Ibaraki, Japan.
Mohamed
A
A
Onodera
T
T
Kato
H
H
Ohashi
T
T
Goitsuka
R
R
Tsujimoto
H
H
Hasegawa
A
A
Furusawa
S
S
Yoshihara
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001681:Biological Assay; D002460:Cell Line; D002634:Chemotaxis, Leukocyte; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D018076:DNA, Complementary; D004285:Dogs; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis* / Q000235:genetics; D007668:Kidney; D008070:Lipopolysaccharides / Q000494:pharmacology; D008198:Lymph Nodes / Q000276:immunology; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000502:physiology; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis*; D017386:Sequence Homology, Amino Acid; D013552:Swine; D014162:Transfection
D017931:DNA Primers; D018076:DNA, Complementary; D016209:Interleukin-8; D008070:Lipopolysaccharides; D011994:Recombinant Proteins