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The Gpd gene was amplified from the genomic DNA of Treponema pallidum and cloned into the appropriate site of pcDNA3.1(+) vector. The expression of pcDNA3.1(+)-Gpd in HeLa cells was tested with Western blotting and technology of immunocytochemistry. New Zealand rabbits were immunized with the eukaryotic expression recombinant pcDNA3.1(+)-Gpd. A fusion protein of Gpd with 4.1 kDa has been effectively expressed in HeLa cells, which were detected by Western blotting and the immunocytochemistry techniques. The New Zealand rabbits were able to elicit the specific antibody after immunization with the nucleic acid vaccine. The antibody titer could reach as high as 1∶1024 after 2 weeks of the third injection; and the splenocytes proliferated evidently due to the Gpd protein stimulation. Both the antibody titer and the splenocytes proliferation were higher substantially than those of controls (P<0.01). All above data will contribute to an experimental basis of further study of the biological function of Gpd protein as well as DNA vaccine for syphilis.
The Gpd gene was amplified from the genomic DNA of Treponema pallidum and cloned into the appropriate site of pcDNA3.1 (+) vector. The expression of pcDNA3.1 (+) - Gpd in HeLa cells was tested with Western blotting and technology of immunocytochemistry . New Zealand rabbits were immunized with the eukaryotic expression recombinant pcDNA3.1 (+) - Gpd. A fusion protein of Gpd with 4.1 kDa has been expressed expressed in HeLa cells, which were detected by Western blotting and the immunocytochemistry techniques. The New Zealand rabbits were able to elicit the specific antibody after immunization with the nucleic acid vaccine. The antibody titer could reach as high as 1:1024 after 2 weeks of the third injection; and the splenocytes proliferated evidently due to the Gpd protein stimulation. Both the antibody titer and the splenocytes proliferation were higher than those of controls (P <0.01). All above data will contribute to an experimental basis of further study of the biological function of Gpd protein as well as DNA vaccine for syphilis.