A gene encoding a cysteine proteinase from Paragonimus westermani has been cloned and expressed in Escherichia coli. The cysteine proteinase cDNA fragment was amplified by reverse transcription–polymerase chain reaction (RT-PCR) using degenerate oligonucleotide primers derived from the conserved active site of the cysteine proteinase. The 5′ and 3′ regions of the gene were amplified using a PCR technique for the rapid amplification of cDNA ends. The cloned gene has an open reading frame of 687 bp and deduced amino acid sequence of 229. Sequence analysis and alignment showed significant homologies with the eukaryotic cysteine proteinases and conservation of the Cys, His, and Asp residues that form a catalytic triad. Analysis of the expressed protein on sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that the molecular weight of the protein was approximately 28.5 kDa. The expressed protein reacted with the sera of patients with paragonimiasis but not with the sera of fascioliasis and clonorchiasis. These results suggest that the expressed protein may be valuable as a specific diagnostic material for the immunodiagnosis of paragonimiasis.
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April 2000
MOLECULAR-CELL BIOLOGY|
April 01 2000
CLONING AND EXPRESSION OF A CYSTEINE PROTEINASE GENE FROM PARAGONIMUS WESTERMANI ADULT WORMS
Tong-Soo Kim;
Tong-Soo Kim
Department of Parasitology, National Institute of Health, Seoul 122-701, Korea
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Byoung-Kuk Na;
Byoung-Kuk Na
Department of Parasitology, National Institute of Health, Seoul 122-701, Korea
*Department of Biology, College of Natural Science, Chung-Ang University, Seoul 156-756, Korea.
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Po-Hyun Park;
Po-Hyun Park
Department of Parasitology, National Institute of Health, Seoul 122-701, Korea
*Department of Biology, College of Natural Science, Chung-Ang University, Seoul 156-756, Korea.
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Kyoung-Ju Song;
Kyoung-Ju Song
Department of Parasitology, National Institute of Health, Seoul 122-701, Korea
*Department of Biology, College of Natural Science, Chung-Ang University, Seoul 156-756, Korea.
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Nicolas Hontzeas;
Nicolas Hontzeas
Department of Parasitology, National Institute of Health, Seoul 122-701, Korea
†Department of Zoology, College of Biological Science, University of Guelph, Guelph, Ontario, Canada N1G 2W1
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Chul-Yong Song
Chul-Yong Song
Department of Parasitology, National Institute of Health, Seoul 122-701, Korea
*Department of Biology, College of Natural Science, Chung-Ang University, Seoul 156-756, Korea.
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J Parasitol (2000) 86 (2): 333–339.
Citation
Tong-Soo Kim, Byoung-Kuk Na, Po-Hyun Park, Kyoung-Ju Song, Nicolas Hontzeas, Chul-Yong Song; CLONING AND EXPRESSION OF A CYSTEINE PROTEINASE GENE FROM PARAGONIMUS WESTERMANI ADULT WORMS. J Parasitol 1 April 2000; 86 (2): 333–339. doi: https://doi.org/10.1645/0022-3395(2000)086[0333:CAEOAC]2.0.CO;2
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