Cloning, characterization, and expression of a novel secretory lipase-like gene from Clonorchis sinensis.
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.
Hu
Fengyu
F
Institute of Infectious Diseases, Guangzhou No. 8 People's Hospital, Guangzhou, People's Republic of China.
Chen
Wanshan
W
Li
Linhua
L
Lu
Yecheng
Y
Song
Weinan
W
Kuang
Yanling
Y
Zhang
Fuchun
F
D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D003001:Cloning, Molecular; D003004:Clonorchis sinensis / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D016384:Consensus Sequence; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D018509:DNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics*; D007150:Immunohistochemistry; D008049:Lipase / Q000737:chemistry / Q000235:genetics*; D008853:Microscopy; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D021382:Protein Sorting Signals; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D021382:Protein Sorting Signals; D008049:Lipase