Cloning and expression analysis of rainbow trout Oncorhynchus mykiss tumour necrosis factor-alpha

Kerry J Laing, Tiehui Wang, Jun Zou, Jason Holland, Suhee Hong, Niels Bols, Ikuo Hirono, Takashi Aoki, Christopher J Secombes

Research output: Contribution to journalArticle

203 Citations (Scopus)

Abstract

A rainbow trout (Oncorhynchus mykiss) gene for tumor necrosis factor (TNF) has been cloned and sequenced. The cDNA contains an open reading frame of 738 nucleotides that translate into a 246 amino-acid putative peptide, with a 5' untranslated region (UTR) of 140 bp and a 3' UTR of 506 bp. Two potential N-linked glycosylation sites exist in the translation. The genomic sequence measures 2007 bp and contains three introns that intercept four coding exons. Expression studies using RT-PCR have shown that the trout TNF gene is constitutively expressed in the gill and kidney of unstimulated fish. Trout TNF expression could be up-regulated by stimulation of isolated head kidney leucocytes with lipopolysaccharide (LPS). Similarly, stimulation of a trout macrophage cell line (RTS11) with LPS resulted in an increased transcript level, as did incubation with recombinant trout interleukin (IL)-1 beta. The optimal timing for induction of TNF expression in trout macrophages was determined using recombinant trout IL-1 beta, where a clear induction was apparent by 2 h and peaked at 4 h. Evidence that this TNF gene is equivalent to mammalian TNF-alpha is discussed.
Original languageEnglish
Pages (from-to)1315-1322
Number of pages8
JournalEuropean Journal of Biochemistry
Volume268
Issue number5
DOIs
Publication statusPublished - Mar 2001

Keywords

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Line
  • Cloning, Molecular
  • Exons
  • Gene Expression Profiling
  • Gene Library
  • Gills
  • Glycosylation
  • Humans
  • Interleukin-1
  • Introns
  • Kidney
  • Lipopolysaccharides
  • Macrophages
  • Molecular Sequence Data
  • Oncorhynchus
  • Open Reading Frames
  • RNA, Messenger
  • Sequence Alignment
  • Transcriptional Activation
  • Tumor Necrosis Factor-alpha
  • Gene structure
  • mRNA expression
  • Rainbow Trout
  • Tumour Necrosis
  • Growth-Factor-Beta
  • Messenger-RNA
  • Molecular-Cloning
  • TNF-alpha
  • Sequence
  • Interleukin-1-Beta
  • Receptor
  • Region
  • Genes
  • Chemokine

Cite this

Cloning and expression analysis of rainbow trout Oncorhynchus mykiss tumour necrosis factor-alpha. / Laing, Kerry J; Wang, Tiehui; Zou, Jun; Holland, Jason; Hong, Suhee; Bols, Niels; Hirono, Ikuo; Aoki, Takashi; Secombes, Christopher J.

In: European Journal of Biochemistry, Vol. 268, No. 5, 03.2001, p. 1315-1322.

Research output: Contribution to journalArticle

Laing, Kerry J ; Wang, Tiehui ; Zou, Jun ; Holland, Jason ; Hong, Suhee ; Bols, Niels ; Hirono, Ikuo ; Aoki, Takashi ; Secombes, Christopher J. / Cloning and expression analysis of rainbow trout Oncorhynchus mykiss tumour necrosis factor-alpha. In: European Journal of Biochemistry. 2001 ; Vol. 268, No. 5. pp. 1315-1322.
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abstract = "A rainbow trout (Oncorhynchus mykiss) gene for tumor necrosis factor (TNF) has been cloned and sequenced. The cDNA contains an open reading frame of 738 nucleotides that translate into a 246 amino-acid putative peptide, with a 5' untranslated region (UTR) of 140 bp and a 3' UTR of 506 bp. Two potential N-linked glycosylation sites exist in the translation. The genomic sequence measures 2007 bp and contains three introns that intercept four coding exons. Expression studies using RT-PCR have shown that the trout TNF gene is constitutively expressed in the gill and kidney of unstimulated fish. Trout TNF expression could be up-regulated by stimulation of isolated head kidney leucocytes with lipopolysaccharide (LPS). Similarly, stimulation of a trout macrophage cell line (RTS11) with LPS resulted in an increased transcript level, as did incubation with recombinant trout interleukin (IL)-1 beta. The optimal timing for induction of TNF expression in trout macrophages was determined using recombinant trout IL-1 beta, where a clear induction was apparent by 2 h and peaked at 4 h. Evidence that this TNF gene is equivalent to mammalian TNF-alpha is discussed.",
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T1 - Cloning and expression analysis of rainbow trout Oncorhynchus mykiss tumour necrosis factor-alpha

AU - Laing, Kerry J

AU - Wang, Tiehui

AU - Zou, Jun

AU - Holland, Jason

AU - Hong, Suhee

AU - Bols, Niels

AU - Hirono, Ikuo

AU - Aoki, Takashi

AU - Secombes, Christopher J

PY - 2001/3

Y1 - 2001/3

N2 - A rainbow trout (Oncorhynchus mykiss) gene for tumor necrosis factor (TNF) has been cloned and sequenced. The cDNA contains an open reading frame of 738 nucleotides that translate into a 246 amino-acid putative peptide, with a 5' untranslated region (UTR) of 140 bp and a 3' UTR of 506 bp. Two potential N-linked glycosylation sites exist in the translation. The genomic sequence measures 2007 bp and contains three introns that intercept four coding exons. Expression studies using RT-PCR have shown that the trout TNF gene is constitutively expressed in the gill and kidney of unstimulated fish. Trout TNF expression could be up-regulated by stimulation of isolated head kidney leucocytes with lipopolysaccharide (LPS). Similarly, stimulation of a trout macrophage cell line (RTS11) with LPS resulted in an increased transcript level, as did incubation with recombinant trout interleukin (IL)-1 beta. The optimal timing for induction of TNF expression in trout macrophages was determined using recombinant trout IL-1 beta, where a clear induction was apparent by 2 h and peaked at 4 h. Evidence that this TNF gene is equivalent to mammalian TNF-alpha is discussed.

AB - A rainbow trout (Oncorhynchus mykiss) gene for tumor necrosis factor (TNF) has been cloned and sequenced. The cDNA contains an open reading frame of 738 nucleotides that translate into a 246 amino-acid putative peptide, with a 5' untranslated region (UTR) of 140 bp and a 3' UTR of 506 bp. Two potential N-linked glycosylation sites exist in the translation. The genomic sequence measures 2007 bp and contains three introns that intercept four coding exons. Expression studies using RT-PCR have shown that the trout TNF gene is constitutively expressed in the gill and kidney of unstimulated fish. Trout TNF expression could be up-regulated by stimulation of isolated head kidney leucocytes with lipopolysaccharide (LPS). Similarly, stimulation of a trout macrophage cell line (RTS11) with LPS resulted in an increased transcript level, as did incubation with recombinant trout interleukin (IL)-1 beta. The optimal timing for induction of TNF expression in trout macrophages was determined using recombinant trout IL-1 beta, where a clear induction was apparent by 2 h and peaked at 4 h. Evidence that this TNF gene is equivalent to mammalian TNF-alpha is discussed.

KW - Amino Acid Sequence

KW - Animals

KW - Base Sequence

KW - Cell Line

KW - Cloning, Molecular

KW - Exons

KW - Gene Expression Profiling

KW - Gene Library

KW - Gills

KW - Glycosylation

KW - Humans

KW - Interleukin-1

KW - Introns

KW - Kidney

KW - Lipopolysaccharides

KW - Macrophages

KW - Molecular Sequence Data

KW - Oncorhynchus

KW - Open Reading Frames

KW - RNA, Messenger

KW - Sequence Alignment

KW - Transcriptional Activation

KW - Tumor Necrosis Factor-alpha

KW - Gene structure

KW - mRNA expression

KW - Rainbow Trout

KW - Tumour Necrosis

KW - Growth-Factor-Beta

KW - Messenger-RNA

KW - Molecular-Cloning

KW - TNF-alpha

KW - Sequence

KW - Interleukin-1-Beta

KW - Receptor

KW - Region

KW - Genes

KW - Chemokine

U2 - 10.1046/j.1432-1327.2001.01996.x

DO - 10.1046/j.1432-1327.2001.01996.x

M3 - Article

VL - 268

SP - 1315

EP - 1322

JO - European Journal of Biochemistry

JF - European Journal of Biochemistry

SN - 0014-2956

IS - 5

ER -