Expression and cross-species reactivity of fatty acid-binding protein of Clonorchis sinensis.
Clonorchis sinensis is a Chinese liver fluke that chronically resides in the biliary tract. The fatty acid-binding protein (FABP) is known to play an important role in the intracellular transport of long-chain fatty acids that are obtained by the fluke from the host. Although FABP has stimulated considerable interest as a vaccine target candidate, the nature of FABP from C. sinensis (CsFABP) remains unclear. In this paper, we describe the cloning and expression of recombinant FABP and immune cross-reaction by Western blot analysis. Sequence analysis revealed that the CsFABP cDNA contained a single open reading frame (ORF) coding for 134 amino acids with an estimated molecular mass of a 15.2 kDa. The DNA sequence of CsFABP cDNA showed significant homology to schistosome cytosolic FABPs, with a 49% amino acid sequence identity and 89% similarity to Schistosoma japonicum. This DNA also showed a high sequence similarity at the amino acid level to S. mansoni (Sm14; 83%) and Fasciola hepatica (80%). The CsFABP cDNA was cloned into expression vector pET28a, expressed in Escherichia coli and the recombinant protein purified by affinity chromatography. The recombinant CsFABP was cross-reacted with sera obtained from patients with fascioliasis and paragonimiasis. These results suggest that CsFABP may be useful as a vaccine for clonorchiasis.
Lee
Ji-Sook
JS
Department of Parasitology and Institute of Tropical Medicine, BK 21 Project for Medical Science, Yonsei University College of Medicine, 134 Shinchondong, Seodaemungu, 120-752 Seoul, Korea.
Yong
Tai-Soon
TS
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D001483:Base Sequence; D015153:Blotting, Western; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D003001:Cloning, Molecular; D003004:Clonorchis sinensis / Q000235:genetics* / Q000378:metabolism*; D003429:Cross Reactions; D018076:DNA, Complementary; D018509:DNA, Helminth / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005210:Fasciola hepatica / Q000235:genetics; D005211:Fascioliasis / Q000276:immunology / Q000469:parasitology; D050556:Fatty Acid-Binding Proteins; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D010237:Paragonimiasis / Q000276:immunology / Q000469:parasitology; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D012549:Schistosoma japonicum / Q000235:genetics; D012550:Schistosoma mansoni / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D000909:Antibodies, Helminth; D002352:Carrier Proteins; D018076:DNA, Complementary; D018509:DNA, Helminth; D050556:Fatty Acid-Binding Proteins; D015801:Helminth Proteins; D011994:Recombinant Proteins