Cloning, Expression And Immunogenicity Study On TSOL16 Gene From Taenia Solium Oncosphere

Total RNA was extracted from Taenia solium oncosphere, the TSOL16 gene specific primers were designed using Oligo software. A 559 bp specific fragment was amplified by RT-PCR and ligated into pMD18-T vector. It was identified by restriction endonuclease analysis, PCR and sequencing that the fragment contained the complete open reading frame (ORF) of the TSOL16 gene. The homologies of the nucleotide sequence and encoding amino acid sequence were 99% and 97% respectively compared with the sequence published in Genebank.The recombinant plasmid pMD18-TSOL16 was digested with EcoR I and Sal I, releasing the TSOL16 gene fragment. The fragment subsequently was inserted into the EcoR I and Xho I sites of the prokaryotic expression vector pGEX-4T-1 and was transformed into E.coli BL21, this construct was also sequenced to ensure fidelity. Expression of the recombinant plasmid was induced by IPTG in E. coli resulting in a high level of protein expression. The expressed products were analyzed by SDS-PAGE and Western-blotting. The results of SDS-PAGE showed that the 40 ku TSOL16 GST fusion was expressed successfully. Western-blotting showed good immunoreactivity of the expressed product. Immunized mice with the expressed product, collected the antisera from the immunized mice and examined by ELISA. The were The results showed that immunization of mice generated antibodies and the antibodies specifically reacted with TSOL16.This study cloned TSOL16 gene from Taenia solium oncosphere, successfully and expressed in prokaryotic. The expressed products confirmed good immunogenicity. It provided theoretic proof and material for developing genetic engineering vaccine.