Cloning and analysis of expression of a gilthead sea bream (Sparus aurata) Mx cDNA

C Tafalla, R Aranguren, C J Secombes, A Figueras, B Novoa

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

In the current work, we have cloned and sequenced the full cDNA for a Mx protein in the gilthead sea bream (Sparus aurata) by RACE PCR. The Mx cDNA of 2182 bp contained an open reading frame of 1857 bp that codes for a protein of 618 aa. Within the coding sequence, characteristic features of Mx proteins were found, such as a tripartite guanosine-5'-triphosphate (GTP)-binding motif (GXXXSGKS/T, DXXG and T/NKXD), the signature of the dynamin family, LPRG(S/K)GIVTR, and a sequence that codes for a leucine zipper at the C-terminal region of the protein.

An RT-PCR was optimised to estimate the level of expression of Mx protein in sea bream. Through this method we determined that Mx is constitutively expressed in head kidney, liver, spleen, heart, gills, muscle and brain of healthy sea bream. Intramuscular challenge of sea bream with polyinosinic:polycytidylic acid (Poly I:C) up-regulated Mx expression in liver, head kidney, spleen and muscle. Constitutive expression was also found in isolated head kidney macrophages and blood leukocytes. This expression was significantly up-regulated by addition of Poly I:C. Mx was not constitutively expressed in the sea bream established cell line, SAF-1, but Poly I:C and nodavirus were also capable of inducing Mx expression in this cell line. (C) 2003 Elsevier Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)11-24
Number of pages14
JournalFish & Shellfish Immunology
Volume16
DOIs
Publication statusPublished - 2004

Keywords

  • Mx
  • sea bream
  • Spurns aurata
  • cytokine
  • macrophages
  • poly I : C
  • SAF-1 cell line
  • nodavirus
  • DOUBLE-STRANDED-RNA
  • PANCREATIC NECROSIS VIRUS
  • SALMON-ANEMIA-VIRUS
  • POLY I-C
  • RAINBOW-TROUT
  • MOLECULAR-CLONING
  • ATLANTIC SALMON
  • INFLUENZA-VIRUS
  • CONFERS RESISTANCE
  • PROTEIN

Cite this

Cloning and analysis of expression of a gilthead sea bream (Sparus aurata) Mx cDNA. / Tafalla, C ; Aranguren, R ; Secombes, C J ; Figueras, A ; Novoa, B .

In: Fish & Shellfish Immunology, Vol. 16, 2004, p. 11-24.

Research output: Contribution to journalArticle

Tafalla, C ; Aranguren, R ; Secombes, C J ; Figueras, A ; Novoa, B . / Cloning and analysis of expression of a gilthead sea bream (Sparus aurata) Mx cDNA. In: Fish & Shellfish Immunology. 2004 ; Vol. 16. pp. 11-24.
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abstract = "In the current work, we have cloned and sequenced the full cDNA for a Mx protein in the gilthead sea bream (Sparus aurata) by RACE PCR. The Mx cDNA of 2182 bp contained an open reading frame of 1857 bp that codes for a protein of 618 aa. Within the coding sequence, characteristic features of Mx proteins were found, such as a tripartite guanosine-5'-triphosphate (GTP)-binding motif (GXXXSGKS/T, DXXG and T/NKXD), the signature of the dynamin family, LPRG(S/K)GIVTR, and a sequence that codes for a leucine zipper at the C-terminal region of the protein.An RT-PCR was optimised to estimate the level of expression of Mx protein in sea bream. Through this method we determined that Mx is constitutively expressed in head kidney, liver, spleen, heart, gills, muscle and brain of healthy sea bream. Intramuscular challenge of sea bream with polyinosinic:polycytidylic acid (Poly I:C) up-regulated Mx expression in liver, head kidney, spleen and muscle. Constitutive expression was also found in isolated head kidney macrophages and blood leukocytes. This expression was significantly up-regulated by addition of Poly I:C. Mx was not constitutively expressed in the sea bream established cell line, SAF-1, but Poly I:C and nodavirus were also capable of inducing Mx expression in this cell line. (C) 2003 Elsevier Ltd. All rights reserved.",
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T1 - Cloning and analysis of expression of a gilthead sea bream (Sparus aurata) Mx cDNA

AU - Tafalla, C

AU - Aranguren, R

AU - Secombes, C J

AU - Figueras, A

AU - Novoa, B

PY - 2004

Y1 - 2004

N2 - In the current work, we have cloned and sequenced the full cDNA for a Mx protein in the gilthead sea bream (Sparus aurata) by RACE PCR. The Mx cDNA of 2182 bp contained an open reading frame of 1857 bp that codes for a protein of 618 aa. Within the coding sequence, characteristic features of Mx proteins were found, such as a tripartite guanosine-5'-triphosphate (GTP)-binding motif (GXXXSGKS/T, DXXG and T/NKXD), the signature of the dynamin family, LPRG(S/K)GIVTR, and a sequence that codes for a leucine zipper at the C-terminal region of the protein.An RT-PCR was optimised to estimate the level of expression of Mx protein in sea bream. Through this method we determined that Mx is constitutively expressed in head kidney, liver, spleen, heart, gills, muscle and brain of healthy sea bream. Intramuscular challenge of sea bream with polyinosinic:polycytidylic acid (Poly I:C) up-regulated Mx expression in liver, head kidney, spleen and muscle. Constitutive expression was also found in isolated head kidney macrophages and blood leukocytes. This expression was significantly up-regulated by addition of Poly I:C. Mx was not constitutively expressed in the sea bream established cell line, SAF-1, but Poly I:C and nodavirus were also capable of inducing Mx expression in this cell line. (C) 2003 Elsevier Ltd. All rights reserved.

AB - In the current work, we have cloned and sequenced the full cDNA for a Mx protein in the gilthead sea bream (Sparus aurata) by RACE PCR. The Mx cDNA of 2182 bp contained an open reading frame of 1857 bp that codes for a protein of 618 aa. Within the coding sequence, characteristic features of Mx proteins were found, such as a tripartite guanosine-5'-triphosphate (GTP)-binding motif (GXXXSGKS/T, DXXG and T/NKXD), the signature of the dynamin family, LPRG(S/K)GIVTR, and a sequence that codes for a leucine zipper at the C-terminal region of the protein.An RT-PCR was optimised to estimate the level of expression of Mx protein in sea bream. Through this method we determined that Mx is constitutively expressed in head kidney, liver, spleen, heart, gills, muscle and brain of healthy sea bream. Intramuscular challenge of sea bream with polyinosinic:polycytidylic acid (Poly I:C) up-regulated Mx expression in liver, head kidney, spleen and muscle. Constitutive expression was also found in isolated head kidney macrophages and blood leukocytes. This expression was significantly up-regulated by addition of Poly I:C. Mx was not constitutively expressed in the sea bream established cell line, SAF-1, but Poly I:C and nodavirus were also capable of inducing Mx expression in this cell line. (C) 2003 Elsevier Ltd. All rights reserved.

KW - Mx

KW - sea bream

KW - Spurns aurata

KW - cytokine

KW - macrophages

KW - poly I : C

KW - SAF-1 cell line

KW - nodavirus

KW - DOUBLE-STRANDED-RNA

KW - PANCREATIC NECROSIS VIRUS

KW - SALMON-ANEMIA-VIRUS

KW - POLY I-C

KW - RAINBOW-TROUT

KW - MOLECULAR-CLONING

KW - ATLANTIC SALMON

KW - INFLUENZA-VIRUS

KW - CONFERS RESISTANCE

KW - PROTEIN

U2 - 10.1016/S1050-4648(03)00010-X

DO - 10.1016/S1050-4648(03)00010-X

M3 - Article

VL - 16

SP - 11

EP - 24

JO - Fish & Shellfish Immunology

JF - Fish & Shellfish Immunology

SN - 1050-4648

ER -