Purification and characterization of a 7-kDa protein from Clonorchis sinensis adult worms.

Lee HJ, Lee CS, Kim BS, Joo KH, Lee JS, Kim TS, Kim HR
J Parasitol. 2002 88 (3): 499-504

PMID: 12099418 · DOI:10.1645/0022-3395(2002)088[0499:PACOAK]2.0.CO;2

A 7-kDa protein was purified from extracts of adult Clonorchis sinensis by a combination of ammonium sulfate precipitation, anion exchange chromatography, cation exchange chromatography, gel-filtration chromatography, and reversed-phase FPLC. The 7-kDa protein exists in the excretory-secretory products of adult C. sinensis, but not in extracts of adult Paragonimus westermani. Also, the 7-kDa protein reacted with the sera of patients with clonorchiasis but not with paragonimiasis or normal human sera. To observe the localization of the 7-kDa protein in the tissue of adult C. sinensis, an immunogold labeling method was followed using anti-7-kDa antibody. The gold particles were observed in the basal layer below the tegumental syncytium, in the interstitial matrix of the parenchyma, and in the content of the uterus. The 7-kDa cDNA was obtained through reverse transcription-polymerase chain reaction using a primer designed from N-terminal sequence analysis. Rapid amplification of cDNA ends (5'-RACE) was used to obtain the complete protein coding sequence. The sequence encodes a 90-amino acid polypeptide. The deduced amino acid sequence of the 7-kDa protein revealed no homology with proteins of different organisms reported so far. These results suggest that the 7-kDa protein is a fluid antigen and may be valuable as a tool for the immunodiagnosis of clonorchiasis.

MeSH Terms (19)

Amino Acid Sequence Animals Antigens, Helminth Base Sequence Blotting, Western Chromatography, Gel Chromatography, Ion Exchange Clonorchiasis Clonorchis sinensis DNA, Complementary Electrophoresis, Polyacrylamide Gel Humans Male Mice Microscopy, Electron Molecular Sequence Data Reverse Transcriptase Polymerase Chain Reaction RNA, Helminth Sequence Analysis, DNA

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