[Expression and immunocompetence characterization of Plasmodium falciparum lactate dehydrogenase].
Wu Ys, Min Li, Dong Wq, Li Yj
Abstract
Wu Ys, Min Li, Dong Wq, Li Yj
Abstract
OBJECTIVE: To express lactate dehydrogenase (LDH) gene of Plasmodium falciparum FCC1/HN in the E. coli TG1 and analyse its immunocompetence. METHODS: The LDH gene of the P. falciparum was specifically amplified by polymerase chain reaction, and the recovered gene fragment was cloned into pGEX-4T-1 vector for expression of fusion protein with glutathione S-transferase(GST). The recombinant plasmid was transformed into the E. coli TG1. Four mice (Kunming strain) were immunized with purified expressed protein(antigen) and the polyclonal antibodies were collected. The immunocompetence of recombinant protein was analysed by ELISA and Western-blot. RESULTS: The LDH gene of P. falciparum was successfully expressed in the E. coli TG1. The expressed protein exhibited a specific reaction with immune sera obtained from rabbits immunized with P. falciparum. The specific humoral responses were induced in mice and the titer of the specific antibody was 1:16 by two-dimensional diffusion assay. CONCLUSION: The LDH gene of P. falciparum has been successfully expressed in the E. coli TG1 and the expressed protein has high antigencity.
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OBJECTIVE: To express lactate dehydrogenase (LDH) gene of Plasmodium falciparum FCC1/HN in the E. coli TG1 and analyse its immunocompetence. METHODS: The LDH gene of the P. falciparum was specifically amplified by polymerase chain reaction, and the recovered gene fragment was cloned into pGEX-4T-1 vector for expression of fusion protein with glutathione S-transferase(GST). The recombinant plasmid was transformed into the E. coli TG1. Four mice (Kunming strain) were immunized with purified expressed protein(antigen) and the polyclonal antibodies were collected. The immunocompetence of recombinant protein was analysed by ELISA and Western-blot. RESULTS: The LDH gene of P. falciparum was successfully expressed in the E. coli TG1. The expressed protein exhibited a specific reaction with immune sera obtained from rabbits immunized with P. falciparum. The specific humoral responses were induced in mice and the titer of the specific antibody was 1:16 by two-dimensional diffusion assay. CONCLUSION: The LDH gene of P. falciparum has been successfully expressed in the E. coli TG1 and the expressed protein has high antigencity.
Key concepts: Polyclonal antibodies, Molecular biology, Recombinant DNA, Biology, Immunocompetence, Plasmodium falciparum, Lactate dehydrogenase, Antibody