The rSjTGR-Sec was expressed and harvested as described previously [13 (link), 19 (link)]. Most of rSjTGR-Sec was expressed in E. coli as a soluble His-tagged fusion protein when bacterial growth occurred at 24 °C. The rSjTGR-Sec was purified by His Mag™ Agarose beads according to manufacturer’s instructions. The concentration of purified rSjTGR-Sec was determined using a Bradford assay kit (Sangon, China), and the protein was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and stored at 4 °C [20 (link)].
The SWAP containing wtSjTGR was prepared from adult schistosomes. Rabbits were infected with cercariae of S. japonicum by exposing their abdominal skin and were sacrificed at 42 days after infection to collect the adult schistosomes by perfusion. The worms were washed three times with phosphate-buffered saline (PBS). After sufficient grinding of the adult schistosomes in PBS, a suitable amount of benzoyl sulfonyl fluoride (PMSF) solution was added to obtain a final concentration of 1 mM. The homogenate was centrifuged at 4 °C, 12,000×g for 20 min [1 (link), 10 (link)]. The concentration of supernatant containing wtSjTGR was determined by Bradford assay kit (Sangon, China).
The SWAP containing wtSjTGR was prepared from adult schistosomes. Rabbits were infected with cercariae of S. japonicum by exposing their abdominal skin and were sacrificed at 42 days after infection to collect the adult schistosomes by perfusion. The worms were washed three times with phosphate-buffered saline (PBS). After sufficient grinding of the adult schistosomes in PBS, a suitable amount of benzoyl sulfonyl fluoride (PMSF) solution was added to obtain a final concentration of 1 mM. The homogenate was centrifuged at 4 °C, 12,000×g for 20 min [1 (link), 10 (link)]. The concentration of supernatant containing wtSjTGR was determined by Bradford assay kit (Sangon, China).
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