title: Cloning and characterization of a TNF-like protein of Plecoglossus altivelis (ayu fish).
abstract: Ayu TNF cDNA contains an open reading frame of 708bp encoding 235 amino acids. Poly adeniration (A) signal and eight AU-rich sequences were present in 858bp 3' UTR. Southern blot analysis indicated that ayu TNF is single-copy gene. The genomic DNA sequence of ayu TNF, consisting of four exons and three introns, was shown to be conserved well throughout evolution from fish to mammals. The amino acid sequence of ayu TNF was shown to have 32-41% of amino acid identity to other known fish TNF, and about 30% of amino acid identity to mammalian TNFs. A phylogenetic analysis based on the amino acid sequence of TNF indicated that ayu has a distinctive evolutionary path. Also, two residues of cysteine important for the formation of the three-dimensional structure were conserved in ayu TNF. For the functional analysis, ayu TNF was inserted into expression vector pCold/TF, transferred into Chaperone Competent Cells BL21 (pKJE7); this produced soluble mature ayu recombinant TNF. Ayu recombinant TNF was shown to induce respiratory burst activity from ayu kidney. The above results indicate that ayu TNF plays an important role in phylaxis, as it does in mammals.
authors.affiliation: Institute for Health and Sciences, Tokushima Bunri University, Nishihama, Yamashiro-cho, Tokushima 770-8514, Japan.
authors.forename: Maya
authors.initials: M
authors.lastname: Uenobe
authors.forename: Chie
authors.initials: C
authors.lastname: Kohchi
authors.forename: Noriko
authors.initials: N
authors.lastname: Yoshioka
authors.forename: Akihiko
authors.initials: A
authors.lastname: Yuasa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Inagawa
authors.forename: Kayoko
authors.initials: K
authors.lastname: Morii
authors.forename: Takashi
authors.initials: T
authors.lastname: Nishizawa
authors.forename: Yukinori
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Gen-Ichiro
authors.initials: G
authors.lastname: Soma
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000737:chemistry / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D006801:Humans; D008969:Molecular Sequence Data; D036441:Osmeriformes / Q000235:genetics*; D016897:Respiratory Burst / Q000276:immunology; D017422:Sequence Analysis, DNA; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry* / Q000235:genetics*
chemical_list: D000890:Anti-Infective Agents; D014409:Tumor Necrosis Factor-alpha