[Cloning, expression and prediction of protein structure of Ts3 gene of Cysticercus cellulosae].

OBJECTIVE

To discover a new gene of candidate molecule for the diagnosis of cysticercosis.

METHODS

Cysticercus cellulosae cDNA library was immunoscreened with pooled serum of cysticercosis patients. The encoded protein of a new gene, Ts3, was analyzed through biology software (EXPASY) on line. The new gene was cloned, the prokaryotic expression vector pET28a (+)-Ts3 was constructed, and the expressed result was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.

RESULTS

The new gene Ts3 (GenBank accession No. EU338456) was obtained with 531 bp and an ORF of C. cellulosae. With relatively stable structure and physicochemical properties, the new gene was anon-transmembrane protein and contained more protein kinase C phosphorylation site. To induce its expression, the Ts3 gene was cloned into the vector pET28a (+), and a protein with a relative molecular mass Mr21000 was obtained which was recognized by the sera of cysticercosis cases.

CONCLUSION

The Ts3 gene of C. cellulosae was expressed, and the protein shows adequate antigenicity.