Peptidases of Trichobilharzia regenti (Schistosomatidae) and its molluscan host Radix peregra S. Lat. (Lymnaeidae): construction and screening of cDNA library from intramolluscan stages of the parasite.
Trichobilharzia regenti is a neurotropic bird schistosome,causing cercarial dermatitis in humans. In this study, ZAP cDNA expression library from Radix peregra s. lat. hepatopancreases containing intramolluscan stages of T. regenti was constructed and screened using PCR with specific and degenerate primers, designed according to previously described serine and cysteine peptidases of other parasite species. Full-length sequences of cathepsins B1 and L, and two serine peptidases, named RpSP1 and RpSP2, were obtained. The protein-protein BLAST analysis and parallel control reactions with template from hepatopancreases of T. regenti non-infected snails revealed that only cathepsin B1 was of parasite origin. The remaining sequences were derived from the snail intermediate host, which implies that the initial source of parasite mRNA was contaminated by snail tissue. Regardless of this contamination, the cDNA library remains an excellent molecular tool for detection and identification of bioactive molecules in T. regenti cercariae.
Dolecková
Katerina
K
Department of Parasitology, Faculty of Science, Charles University in Prague, Vinicná 7, 128 44 Prague 2, Czech Republic. k.doleckova@centrum.cz
Kasný
Martin
M
Mikes
Libor
L
Mutapi
Francisca
F
Stack
Colin
C
Mountford
Adrian P
AP
Horák
Petr
P
D000818:Animals; D002401:Cathepsin B / Q000235:genetics; D056668:Cathepsin L; D002403:Cathepsins / Q000235:genetics; D003546:Cysteine Endopeptidases / Q000235:genetics; D015723:Gene Library; D015801:Helminth Proteins / Q000235:genetics; D008018:Life Cycle Stages; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012551:Schistosomatidae / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012908:Snails / Q000469:parasitology*
D015801:Helminth Proteins; D002403:Cathepsins; D010447:Peptide Hydrolases; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; C534281:CTSL protein, human; D056668:Cathepsin L