Expression and Immunological Character Research of Three Kinds of Food-poisoning Vibrio Poly-recombinant Toxin Gene |
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| Authors: | LI Tao XU Wen-chao SU Ya-bin MA Fei CUI Yi-jie LI Hua-feng |
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| Institution: | Tangshan Entry-exit Inspection and Quarantine Bureau, Tangshan 063000, China |
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| Abstract: | In this study,tdh-vvhA-ctB was constructed using the flexible Linker sequence (Gly4Ser) in order to construct three kinds of food-poisoning Vibrio poly-recombinant toxin gene and recombinant expression vector. The recombinant expression plasmid pET-22b(+)-TVC was constructed and expressed in prokaryotic expression vector. The animals were immunized using the expressed protein after purification to get serum antibody. The specificity and sensitivity of the antibody were verified by agar diffusion test and enzyme-linked immunosorbent assay (ELISA).The results showed that the recombinant expressing plasmid pET-22b(+)-TVC was constructed and expressed successfully in prokaryotic expression vector, the expression level was 11.38%. The expressed protein was mainly inclusion body and a small amount of soluble protein. The gene length was 2 196 bp, encoding 731 amino acids with molecular weight of 81.7 ku. The results were 99.6% homologous to the designed sequence. Agar diffusion reaction and ELISA tests indicated the poly-recombinant toxin TVC could product different immune intersect reaction with other food-poisoning Vibrios but not react with some no-objective bacteria. Expression plasmid of poly-recombinant toxin gene was constructed and serum antibody was prepared successfully. It might be used to check the objective toxin based on the poly-recombinant toxin gene and established the board-spectrum, quick, special detecting way to lay the theoretical and technical basis. |
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| Keywords: | food-poisoning Vibrios poly-recombinant toxin expression immunological character |
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