Analysis of the genes expressed in Clonorchis sinensis adults using the expressed sequence tag approach.

Clonorchis sinensis is a biliary tract parasite, which infects over 30 million people in China, Korea and Southeast Asia through the ingestion of undercooked freshwater fish that harbour the infective metacercariae. The genes expressed in C. sinensis adults were identified in order to develop novel drugs, better diagnostics and vaccines for the parasite. The C. sinensis cDNA library was constructed and DNA sequencing was performed with 450 randomly selected clones. Four hundred and fifteen clones contained the amino-acid-encoding sequences. The functions of these genes could be assigned by DNA sequence homology. Basic Local Alignment Search Tool X analysis showed that 277 of the 415 clones were strongly matched ( P<10(-9)) to previously identified proteins, while the remaining 138 fell into the "no database match" category. Among the clones matching previously identified proteins, 220 putatively identified genes were sorted into seven functional categories. These included the genes associated with energy metabolism (38), gene expression/RNA metabolism (21), regulatory/signalling components (14), protein metabolism/sorting (98), the structure/cytoskeleton (29), membrane transporters (ten) and antigenic proteins (ten). The remaining 57 clones were not included in these categories. The dataset included the genes encoding the proteases, a lipid binding protein, the antigen proteins and the other genes of interest from a diagnostics, drug or vaccine development viewpoint. The present expressed sequence tag analysis proved to be an effective tool for examining gene expression and identified several important genes which increase and complement our knowledge of the biology of C. sinensis.