Abstract:

To obtain the 14-3-3 protein of Trichinella spiralis (T1) and to identify and evaluate its biological characteristics, the biological information of 14-3-3(Ts14-3-3) protein of Trichinella spiralis was analyzed and predicted using relevant websites and software. Ts14-3-3 was recombined with pET-28a using gene cloning and prokaryotic expression techniques, and the recombinant Ts14-3-3(rTs14-3-3) protein was obtained. After the antigenicity was identified by Western blot, the rTs14-3-3 protein immune serum was prepared. qPCR and indirect immunofluorescence were used to identify its expression in different insect stages and its localization in muscle larvae. The effect of rTs14-3-3 protein on larvae in the muscle of mice infected with Trichinella spiralis was evaluated by animal experiments. Soluble rTs14-3-3 protein was successfully prepared, with a size of about 33 kDa, and it could specifically recognize the positive serum of porcine Trichinella spiralis. The titer of the prepared immune serum was as high as 1:52000. The protein was highly expressed in the muscle larvae of Trichinella spiralis, and the gene was transcribed in the larvae, adults, and muscle larvae during the intestinal infection stage of Trichinella spiralis. The pest reduction rate of mouse muscle larvae was 55.2%. rTs14-3-3 protein has good reactivity and immunogenicity, can reduce the number of muscle larvae in mouse tissues infected by Trichinella spiralis, and has potential as a serological diagnosis and candidate vaccine molecular antigen.

Key words: Trichinella spiralis, Trichinella spiralis 14-3-3 protein, bioinformatics, prokaryotic expression, antigenic analysis, immune protection