Bioinformatics analysis on two new genes of Trichinella spiralis and their expression in E.coli system

Objective To predict the antigenicity of two new Trichinella genes,Ad48 h-Ts3 and tetraspanin,synthesize their gene sequences,express their recombinant proteins and evaluate their potential for diagnosis.Methods The potential antigenicity of the two proteins,tetraspanin family protein and adult stage protein Ad48 h-Ts3 were determined according to the result of their hydrophilicity and hydrophobicity predictions.The sequences of these two genes were synthesized after codon optimization and were subcloned into pET28a expression vector and expressed in E.coli system.Western blotting was performed to evaluate the diagnosis value of the proteins using mouse infected sera.Results Bioninformatic analysis showed that tetraspanin gene of Trichinella spiralis,which bears two transmembrane domains,has homology gene in other species.However,Ad48 h-Ts3 gene,which has plenty of antigenic epitopes,is unique in Trichinella spiralis.Genes were subcloned into expression vector and recombinant Ad48 h-Ts3 protein was expressed successfully in E.coli system while no obvious expression was observed in tetraspaning expression.Western blotting showed the expressive recombinant Ad48 hTs3 protein could be recognized by Trichinella infected sera obtained from mice.Conclusion The adult stage protein,Ad48 h-Ts3 protein was expressed successfully in E.coli and had a potential value for diagnosis. Key words: Trichinella spiralis;  Diagnosis;  Tetraspain;  Adult stage protein