Molecular cloning, characterization, and expression of TNF cDNA and gene from Japanese flounder Paralychthys olivaceus

I. Hirono, B. H. Nam, Tomofumi Kurobe, T. Aoki

Research output: Contribution to journalArticle

181 Citations (Scopus)

Abstract

We cloned a cDNA and the gene for Japanese flounder TNF. The TNF cDNA consisted of 1217 bp, which encoded 225 amino acid residues. The identities between Japanese flounder TNF and members of the mammalian TNF family were ~20-30%. The positions of cysteine residues that are important for disulfide bonds were conserved with respect to those in mammalian TNF-α. The Japanese flounder TNF gene has a length of ~2 kbp and consists of four exons and three introns. The positions of the exon-intron junction positions of Japanese flounder TNF gene are similar to those of human TNF-α. However, the length of the first intron of Japanese flounder is much shorter than that of the human TNF-α gene. There are simple CA or AT dinucleotide repeats in the 5'-upstream and 3'-downstream regions of the Japanese flounder TNF gene. Southern blot hybridization indicted that Japanese flounder TNF exists as a single copy. Expression of Japanese flounder TNF mRNA is greatly induced after stimulation of PBLs with LPS, Con A, or PMA. These results indicated that Japanese flounder TNF is more like mammalian TNF-α than mammalian lymphotoxin-α, with respect to its gene structure, length of amino acid sequence, number and position of cysteine residues, and regulation of gene expression.

Original languageEnglish (US)
Pages (from-to)4423-4427
Number of pages5
JournalJournal of Immunology
Volume165
Issue number8
DOIs
StatePublished - Oct 15 2000
Externally publishedYes

Fingerprint

Flounder
Molecular Cloning
Complementary DNA
Genes
Introns
Cysteine
Exons
Dinucleotide Repeats
Lymphotoxin-alpha
Gene Expression Regulation
Southern Blotting
Disulfides
Amino Acid Sequence
Amino Acids
Messenger RNA

ASJC Scopus subject areas

  • Immunology

Cite this

Molecular cloning, characterization, and expression of TNF cDNA and gene from Japanese flounder Paralychthys olivaceus. / Hirono, I.; Nam, B. H.; Kurobe, Tomofumi; Aoki, T.

In: Journal of Immunology, Vol. 165, No. 8, 15.10.2000, p. 4423-4427.

Research output: Contribution to journalArticle

@article{7a8ab63a32754a568eef21cc7deb7df1,
title = "Molecular cloning, characterization, and expression of TNF cDNA and gene from Japanese flounder Paralychthys olivaceus",
abstract = "We cloned a cDNA and the gene for Japanese flounder TNF. The TNF cDNA consisted of 1217 bp, which encoded 225 amino acid residues. The identities between Japanese flounder TNF and members of the mammalian TNF family were ~20-30{\%}. The positions of cysteine residues that are important for disulfide bonds were conserved with respect to those in mammalian TNF-α. The Japanese flounder TNF gene has a length of ~2 kbp and consists of four exons and three introns. The positions of the exon-intron junction positions of Japanese flounder TNF gene are similar to those of human TNF-α. However, the length of the first intron of Japanese flounder is much shorter than that of the human TNF-α gene. There are simple CA or AT dinucleotide repeats in the 5'-upstream and 3'-downstream regions of the Japanese flounder TNF gene. Southern blot hybridization indicted that Japanese flounder TNF exists as a single copy. Expression of Japanese flounder TNF mRNA is greatly induced after stimulation of PBLs with LPS, Con A, or PMA. These results indicated that Japanese flounder TNF is more like mammalian TNF-α than mammalian lymphotoxin-α, with respect to its gene structure, length of amino acid sequence, number and position of cysteine residues, and regulation of gene expression.",
author = "I. Hirono and Nam, {B. H.} and Tomofumi Kurobe and T. Aoki",
year = "2000",
month = "10",
day = "15",
doi = "10.4049/jimmunol.165.8.4423",
language = "English (US)",
volume = "165",
pages = "4423--4427",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "8",

}

TY - JOUR

T1 - Molecular cloning, characterization, and expression of TNF cDNA and gene from Japanese flounder Paralychthys olivaceus

AU - Hirono, I.

AU - Nam, B. H.

AU - Kurobe, Tomofumi

AU - Aoki, T.

PY - 2000/10/15

Y1 - 2000/10/15

N2 - We cloned a cDNA and the gene for Japanese flounder TNF. The TNF cDNA consisted of 1217 bp, which encoded 225 amino acid residues. The identities between Japanese flounder TNF and members of the mammalian TNF family were ~20-30%. The positions of cysteine residues that are important for disulfide bonds were conserved with respect to those in mammalian TNF-α. The Japanese flounder TNF gene has a length of ~2 kbp and consists of four exons and three introns. The positions of the exon-intron junction positions of Japanese flounder TNF gene are similar to those of human TNF-α. However, the length of the first intron of Japanese flounder is much shorter than that of the human TNF-α gene. There are simple CA or AT dinucleotide repeats in the 5'-upstream and 3'-downstream regions of the Japanese flounder TNF gene. Southern blot hybridization indicted that Japanese flounder TNF exists as a single copy. Expression of Japanese flounder TNF mRNA is greatly induced after stimulation of PBLs with LPS, Con A, or PMA. These results indicated that Japanese flounder TNF is more like mammalian TNF-α than mammalian lymphotoxin-α, with respect to its gene structure, length of amino acid sequence, number and position of cysteine residues, and regulation of gene expression.

AB - We cloned a cDNA and the gene for Japanese flounder TNF. The TNF cDNA consisted of 1217 bp, which encoded 225 amino acid residues. The identities between Japanese flounder TNF and members of the mammalian TNF family were ~20-30%. The positions of cysteine residues that are important for disulfide bonds were conserved with respect to those in mammalian TNF-α. The Japanese flounder TNF gene has a length of ~2 kbp and consists of four exons and three introns. The positions of the exon-intron junction positions of Japanese flounder TNF gene are similar to those of human TNF-α. However, the length of the first intron of Japanese flounder is much shorter than that of the human TNF-α gene. There are simple CA or AT dinucleotide repeats in the 5'-upstream and 3'-downstream regions of the Japanese flounder TNF gene. Southern blot hybridization indicted that Japanese flounder TNF exists as a single copy. Expression of Japanese flounder TNF mRNA is greatly induced after stimulation of PBLs with LPS, Con A, or PMA. These results indicated that Japanese flounder TNF is more like mammalian TNF-α than mammalian lymphotoxin-α, with respect to its gene structure, length of amino acid sequence, number and position of cysteine residues, and regulation of gene expression.

UR - http://www.scopus.com/inward/record.url?scp=0034668013&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034668013&partnerID=8YFLogxK

U2 - 10.4049/jimmunol.165.8.4423

DO - 10.4049/jimmunol.165.8.4423

M3 - Article

VL - 165

SP - 4423

EP - 4427

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 8

ER -