OBJECTIVETo further study the therapy of wasting muscle by myostatin as a new targets, the eucaryotic expression vector coupled the foreign T-helper epitope of tetanus toxin (TT) to the N terminus of myostatin was constructed, and the effects of the gene vaccine on forelimb grip were tested in immunized mice.

METHODSA DNA fragment encoding the TT epitope followed by the N terminus of mature myostatin (330bp) was synthesized. The eucaryotic expression vector of myostatin was constructed and the chinese hamster ovary (CHO) cells were infected with the recombinant plasmids pVAC-TT-Ms by liposome transfection according to routine laboratory procedure. The myostatin expression was tested by cell immunofluorescence technique in transfected CHO. The forelimbs grip were tested in immunized mice with myostatin gene vaccine.

RESULTSThe eucaryotic expression vector of myostatin coupled TT epitope was constructed successfully through the restriction analysis and sequencing. The recombinant plasmids pVAC-TT-Ms met quality criterion as gene vaccine by analysis OD260/280 and electrophoresis. The myostatin expression was detected obviously in transfected CHO. The forelimb grip in immunized mice had an obvious increase. The average value of forelimb grip of the mice immunized with pVAC-TT-Ms was about 29.88% greater than that of control mice.

CONCLUSIONThe construction of eucaryotic expression vector of myostatin coupled TT epitope is successful in expression for recombinant human mature peptide of myostatin. The gene vaccine of myostatin meet quality criterion. The immunized mice has an obvious increase in forelimb grip.