| Summary: | 碩士 === 國立屏東科技大學 === 獸醫學系所 === 98 === Melioidosis caused by Burkholderia pseudomallei ( formerly known as Pseudomonoas psueodmallei ). It is a very important zoonosis. The flagellin protein is highly conserved and contained higher antigenicity in comparison to nucleic acid and amino acid sequences of other gene sequences. In this study the full-length flagellin gene ( fli C ) of Melioidosis was amplified by PCR, and cloned into E. coli expression vectors for recombinant flagellin expression. Tnen immunized BALB/c mice with recombinant flagellin protein. The conformation and antigenicity of recombinant flagellin protein were proved by Western-blot. The myeloma cell ( NS-1 ) will fuse with spleen cell from immunized BALB/c mice to produce hybridoma cells and monoclonal antibodies, and subsequently ELISA kit development good sensitivity and specialty. In addition, evaluate the possibility of recombinant flagellin protein for subunit vaccine development. Immunized two times recombinant flagellin protein with 50 μg and 100 μg ,respectively with BALB/c mice and challenged B. pseudomallei. Mice vaccinated with 50 μg recombinant flagellin revealed 50% survived rate,vaccinated with 100 μg recombinant flagellin revealed 75% survived rate. All the mice died in non-immuned group. Thus recombinant flagellin could induce the protective antibodies against B. pseudomallei. In conclusion, the preliminary results demonstrated that the flagellin protein could be a applicant for development of diagnostic kits and applied to detect for B. pseudomallei in Taiwan. The results indicated that recombinant flagellin protein were promising candidate to develop subunit vaccines.
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