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mansoni TGR was amplified by PCR (forward primer 5-CTATTTGGCTAGACGTCTGT-3 and reverse primer 5-AATACAGTTTCCTTCCCGTT-3) and cloned into PCRII-TOPO vector (Invitrogen, The resulting construct was linearized using XhoI (for SP6 RNA polymerase transcription) and SacI (for T7 RNA polymerase transcription)