Clonging, Exprission And Immunogenicity Analysis Of 45W-4B And TSOL18 Genes From Taenia Solium Oncosphere

Taenia solium eggs were collected and total RNA was extracted from hatched and activated eggs in vitro. According to predictive results, the truncated 45W-4B with removal of a signal peptide at N terminal and 19 hydrophobic amino acids at C terminal was amplified by PCR in order that stability and dissolubility of recombinant protein would be improve, the amino acids sequence of 45W-4B and TSOL18 Genes showed 99.7% and 99.5% identity between sequences that we cloned and that reported in GenBank. The PCR products were cloned into the expressing plasmid vector pGEX-4T-1. They were expressed in E. coli as fussion protein with glutathione transferase protein, and were identified by SDS-PAGE, and specific protein band of 40,000 and 42,000 were found, amounting to 35% and 25% of the total proteins of the induced recombinant bacteria at the presence of IPTG Western blotting showed good immunogenicity of the expressed product.Purified recombinant antigens and cysticercus cellulosae coarse extract with 206 adjuvant were made vaccines and immunized pigs in 2 trails. Data suggested that antibody level obviously rose at 15 days and reached peak 50 days after immunization. MTT analysis indicated that proliferation of lymphocyte peaked at 10th days after challenge. In the animal experiments, the parasite decrease rate of 45W-4B and TSOL18 antigens were made 95%and 94%, and in comparison with the cysticercus cellulosae coarse extract did not statistically significant difference, recombinant antigens both were stored at 4℃ for three months, indicated the parasite decrease rate of TSOL18 antigens decreased 51percent, and 45W-4B was up to 97%. TSOL18 and 45W-4B genes were cloned and were expressed, and their protective efficacy was confirmed.