[Cloning, expression and immunologic identification of Eg10 gene of Echinococcus granulosus].

OBJECTIVE

To clone and express Eg10 gene of Echinococcus granulosus, and investigate the immunological characteristic of the recombinant.

METHODS

Eg10 gene was subcloned into pET28a vector. The recombinant plasmid was transformed into E. coli BL21 and induced with IPTG. The recombinant protein was purified with His-bind purification kit. Forty-eight mice were randomly divided into 4 groups. Mice in groups A and B were injected with PBS and PBS+Freund's adjuvant (100 microl) as control. Mice in groups C and D were immunized with 10 mg and 50 microg purified Eg10 antigen formulated in Freund's adjuvant, respectively. All the mice received three immunizations at 2-week intervals with the same dose of antigen. Serum samples were collected at pre-immunization and certain time after immunization. The immunological characteristics of recombinant Eg10 was analyzed by Western blotting and ELISA.

RESULTS

The recombinant Eg10 protein (Mr 31 000) was expressed in E. coli BL21. The recombinant Eg10 and expression product of PET28a/Eg10 were recognized by sera from mice immunized with recombinant Eg10. ELISA showed that the titer of IgG reached a peak at the 8th week in groups C and D, the level of IgG in sera of groups C or D was higher than that of groups A or B (P < 0.05) at the 2nd, 4th, 6th, 8th, and 10th week. There was no significant difference in the level of IgG between group C and group D (P > 0.05).

CONCLUSION

The Eg10 gene has been expressed with immunogenicity.