In vivo expression in skeletal muscle of mice vaccinated with recombinant plasmid pcDNA3-EBA175/HRP II by intramuscular injection.

To observe the in vivo expression of recombinant plasmid pcDNA3 EBA175/HRPⅡ of FCC 1/HN strain Plasmodium falciparum(P.f) which injected into BALB/c mice by intramuscular route. Group 1(experimental group) injected recombinant plasmid 100 μg at left back leg, group 2 (experimental control group) injected blank plasmid pcDNA3 100 μg, group 3(normal control group) injected PBS(pH7.4) 100 μl, and enhanced every 20 days respectively. Taking two sides quadriceps muscles of back leg detected with immunohistochemistry at the 20th day after the 1st injection and at the 10th day after the 2nd injection. Taking the skeletal muscle, heart, liver, lung, kidney and brain at the 40th day after the 3rd injection to draw genomic DNA and detect aim genes by PCR. The results showed that (1) the injected muscle of group 1 was reacted with sera of mice vaccinated with recombinant plasmid, sera of mice immunized with antigen of P.f , 9 peptides HRPⅡMcAb of P.f and McAbs to the whole parasite of P.f respectively, then IgG binding, DAB dyeing, there were brown color pellets on the surface of muscle membrane and in the intramuscular; but which have been observed on or in the muscle of no injecting leg of group 1 and both sides of group 2 and group 3; (2)detecting EBA 175 and HRPⅡgenes by PCR, there were positive results in skeletal muscle, heart, liver, lung and kidney of group 1, and negative results in the same organs and tissues of group 2 and group 3. This study has provided the basis of in vivo expression for developing malaria mixed gene DNA vaccine.