Preparation and Immunological Characteristics of Broad-Spectrum Antibodies Against Poly-Recombinant Toxins from Food-Borne Pathogenic Bacteria.

摘要

Two recombinant toxin genes (HVSVS: Hc1-VT1B-SEA-VT2B-SEB1;VSVHS: VT1B-SEA-VT2B-Hc2-SEB2) of five toxin genes from three genera of bacteria, including E. coli O157:H7, Staphylococcus aureus, and Clostridium botulinum, were constructed by SOE PCR with two linkers encoding five amino acids (GlyProGlyProGly i.e. GPGPG or Gly4Ser). The length of HVSVS gene fragment was 2,937 bp encoding 979 amino acids, and VSVHS was 2,346 bp encoding 782 amino acids. Two recombinant proteins including insoluble protein in pET-22b and inclusion body in pET-28a plasmids were obtained, respectively, and the molecular weights of recombinant fusion proteins were 112.369 kD and 88.899 kD, respectively. Serum antibodies against the five toxins were obtained from rabbits immunized with expressed products. The broad-spectrum antibodies showed high reactive specificity and sensitivity for the five toxins, as well as cross-reactivity with closely related toxins at the same time. These findings provide a foundation for the construction of fusion genes with linkers and for the establishment of broad-spectrum rapid screening methods for food-borne pathogenic bacteria or toxins.