Cloning, characterization, and expression of a novel secretory lipase-like gene from Clonorchis sinensis

Parasitol Res. 2009 Nov;105(6):1661-6. doi: 10.1007/s00436-009-1607-6. Epub 2009 Sep 16.

Abstract

A secretory lipase-like gene was isolated from total cDNA of adult Clonorchis sinensis. The gene has an open reading frame of 1,218 bp long and encodes for a protein of 406 amino acids including a putative signal peptide of 20 amino acids. The deduced amino acid sequence including signal peptide has 42-45% identity with lipase of other species and two typical enzymic active sites that contain consensus sequence (Gly-X-Ser-X-Gly) of lipase. The cDNA encoding this protein was subcloned into pET-28a (+) expression vector and expressed in Escherichia coli. The expressed fusion protein has a molecular mass of about 45 kDa. Prediction of signal peptide and Western blot analysis indicated that the secretory lipase-like protein is an excretory-secretory product of C. sinensis. Immunostaining revealed that the secretory lipase-like protein was localized in the tegument of the adult worm and metacercaria. These results provide basis for further studies on the nutrition taking and invasion of C. sinensis mediated by the secretory lipase-like protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Catalytic Domain
  • Cloning, Molecular
  • Clonorchis sinensis / chemistry
  • Clonorchis sinensis / enzymology*
  • Clonorchis sinensis / genetics
  • Consensus Sequence
  • DNA, Complementary / genetics
  • DNA, Complementary / isolation & purification
  • DNA, Helminth / genetics
  • DNA, Helminth / isolation & purification
  • Escherichia coli / genetics
  • Gene Expression
  • Helminth Proteins / chemistry
  • Helminth Proteins / genetics*
  • Immunohistochemistry
  • Lipase / chemistry
  • Lipase / genetics*
  • Microscopy
  • Molecular Sequence Data
  • Molecular Weight
  • Open Reading Frames
  • Protein Sorting Signals
  • Sequence Alignment
  • Sequence Homology, Amino Acid

Substances

  • DNA, Complementary
  • DNA, Helminth
  • Helminth Proteins
  • Protein Sorting Signals
  • Lipase