Thioredoxin from the silkworm, Bombyx mori: cDNA sequence, expression, and functional characterization

Comp Biochem Physiol B Biochem Mol Biol. 2007 Jul;147(3):574-81. doi: 10.1016/j.cbpb.2007.03.012. Epub 2007 Mar 31.

Abstract

A thioredoxin (Trx) gene was cloned from the silkworm, Bombyx mori. The B. mori Trx (BmTrx) cDNA contains an open reading frame of 318 bp encoding 106 amino acid residues with a conserved active site (CGPC). Northern blot analysis revealed the presence of BmTrx transcripts in all tissues examined. The cDNA encoding BmTrx was expressed as a 12-kDa polypeptide in baculovirus-infected insect Sf9 cells. The recombinant BmTrx proved to be biologically active, using an insulin reduction assay, and was also able to activate thioredoxin peroxidase from B. mori. When H2O2 or paraquat was injected into the body cavity of B. mori larvae, BmTrx mRNA expression was upregulated in the fat body tissue. In addition, the expression levels of BmTrx mRNA in the fat body were greatly increased when B. mori larvae were exposed to low or high temperatures, or injected with microorganisms. These results suggest that BmTrx possibly protects against oxidative stress caused by extreme temperatures and microbial infection as well as by intracellularly generated reactive oxygen species during metabolism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bombyx / genetics*
  • Bombyx / metabolism
  • DNA, Complementary / genetics
  • Enzyme Activation / drug effects
  • Enzyme Activation / physiology
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / physiology*
  • Herbicides / pharmacology
  • Hydrogen Peroxide / pharmacology
  • Insect Proteins / biosynthesis
  • Insect Proteins / genetics*
  • Organ Specificity / drug effects
  • Organ Specificity / physiology
  • Oxidants / pharmacology
  • Oxidative Stress / drug effects
  • Oxidative Stress / physiology
  • Paraquat / pharmacology
  • Peroxidases / metabolism
  • Peroxiredoxins
  • Reactive Oxygen Species / metabolism
  • Thioredoxins / biosynthesis
  • Thioredoxins / genetics*

Substances

  • DNA, Complementary
  • Herbicides
  • Insect Proteins
  • Oxidants
  • Reactive Oxygen Species
  • Thioredoxins
  • Hydrogen Peroxide
  • Peroxidases
  • Peroxiredoxins
  • Paraquat