Antigenic Analysis Of Small Heat Shock Protein Of Cysticercus Cellulosae

A gene encoding small heat-shock protein (sHSP) from Cysticercus cellulosae was amplified by reverse transcript polymerase chain reaction (RT-PCR) technique. The amino acids sequence showed 99.4% identity between sequences that we cloned and that reported in GenBank. The PCR product was cloned into the expressing plasmid vector pGEX-4T-1. It was expressed in E. coli as a fussion protein with glutathione transferase protein, and was indentified by SDS-PAGE, and a specific protein band of 64,000 was found, amounting to 30% of the total proteins of the induced recombinant bacteria at the presence of IPTG. Western blotting and ELISA showed good immunoreactivity of the expressed product. The specific band of expression was excised from the gel and used to immunize mice. The antisera were collected from the immunized mice and examined by ELISA and Western Blotting. The results showed that immunization of mice generated antibodies and the antibodies specifically reacted with recombinant sHSP. It was concluded from these results that the expressed protein of sHSP gene as a fusion protein has antigenicity. From the results of predication sHSP of Cysticercus cellulosae may be a glycol-protein. In the processing of sHSP, the phosphorylated modification may be important. These results suggest that its function may be complicated, related with the regulation of gene expression and maintance of the cellular cytoskeleton and so on.