Objective: To amplify and express the gene from Taenia solium oncosphere in Pichia pastoris system.
Methods: Total RNA was extracted from hatched and activated oncospheres. A 459bp specific fragment was amplified by RT-PCR. It was subcloned into the secreted expression vector pPIC9K, and identified by sequencing and then transformed into P. pastoris SMD1168 by electroporation. PCR analysis showed that 45WB2 was integrated into the genomic DNA of yeast. The recombinants were induced by 1% methanol and the culture supernatant was collected and tested by SDS-PAGE and Western blotting.
Results: 45WB2 gene was expressed successfully in P. pastoris and the product was recognized by human positive serum against cysticercosis.
Conclusion: The recombinant 45WB2 expressed in P. pastoris may be used to prevent pigs from the infection of Taenia solium.