›› 2013, Vol. 31 ›› Issue (3): 8-198-200,205.

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Position Analysis of Three Recombinant Proteins of Echinococcus granulosus Protoscolex with Two-dimensional Electrophoresis

ZHU Ming-xing1,2,3, LI Jun-liang1,2, JU Yan1,2,GAO Peng1,2, ZHAO Wei1,2 *   

  1. 1 Research Center of Medical Science and Technology of Ningxia Medical University, Yinchuan 750004, China; 2 The medical scientific institute of Ningxia, Yinchuan 750004, China; 3 The Experimental Animal Center of Ningxia Medical University, Yinchuan 750004, China
  • Online:2013-06-30 Published:2013-07-17

Abstract:  Objective  To obtain specific antibodies of the three recombinant antigens obtained previously, rEgZW-5, rEg14-3-3 and rEgP-29, for identifiying the corresponding proteins in two-dimensional electrophoretogram of Echinococcus granulosus protoscolex.  Methods  The distribution of proteins from E. granulosus protoscoleces was judged by SDS-PAGE previously. Two-dimensional electrophoresis was used to separate proteins from E. granulosus protoscoleces, and the result was scanned and analyzed by the PDquest software to get the information about the quantity of proteins as well as their isoelectric point (IP) and relative molecular mass (Mr). Rabbits were immunized with the 3 recombinant antigens and antibodies were purified from antisera. Western blotting was used to identify the protein  as marker in two-dimensional electrophoretogram of protoscolex.  Results  SDS-PAGE displayed that the proteins separated from Echinococcus granulosus protoscoleces mainly distributed in the Mr region of 18 000-90 000. 240 proteins were obtained by two-dimensional electrophoresis with Mr 15 790-117 050 and IP 4.0-9.5, and 85.8% (206/241) of the proteins showed the IP ranged from 5 to 9. Western blotting showed that the specific antibody of rEg14-3-3 identified the 14-3-3 protein in two-dimensional electrophoretogram of protoscolex with Mr 33 000 and IP 4.86, the specific antibody of rEgZW-5 identified the ZW-5 protein with Mr 23 000 and IP 4.98, and the specific antibody of rEg P-29 identified the P-29 protein with Mr 29 000 and IP 5.65.  Conclusion  The antibodies against the three recombinant proteins from Echinococcus granulosus protoscoleces can identify corresponding proteins in the two-dimensional electrophotoregrams.

Key words: Echinococcus granulosus, Protoscolex, Recombinant protein, Two-dimensional electrophoresis