RESUMEN
OBJECTIVE To clone and express toxin C (tcdC) gene pathogenicity locus of Clostridium difficile (CD), and prepare the polyclonal antibody against TcdC protein. METHODS The tcdC gene was amplified from CD (ATCC43255) strain genome DNA and inserted into prokaryotic expression vectors. The correct recombinant plasmids were transformed into E.coli which was induced to express the GST-TcdC/L and IL1-TcdC/L fusion protein. Meanwhile, the fusion proteins were respectively purified through Ni-NTA agarose affinity chromatography and Q-Sepharose Fast Flow. The purified GST-TcdC/L fusion protein was used as an antigen to inoculate rabbits to produce antiserum. Two weeks after the final immunization, the rabbits were sacrificed and serum was collected. The titer of the serum was deter-mined by ELISA and the reactivity of the polyclonal antibody was identified by Western blotting. RESULTS The SDS-PAGE result showed that GST-TcdC/L and IL1-TcdC/L fusion proteins were expressed at the size of 45 ku and 39 ku. The titer of TcdC/L polyclonal antibody was over 1:6.4 × 104. Western blotting detection demonstrated that the TcdC/L polyclonal antibody recognized the TcdC protein in CD (ATCC43255). CONCLUSION The tcdC/L gene is cloned and the polyclonal antibody against TcdC/L is prepared, which will contribute to studing the function and mechanism of tcdC gene in pathogenicity locus.
RESUMEN
Botulismo é uma enfermidade que provoca uma paralisia fatal, causada pela toxina do Clostridium botulinum. O botulismo é observado na maioria das espécies avícolas, exceto os abutres, os quais são resistentes. A doença manifesta-se nas aves, apresentando os seguintes sinais clínicos: pescoço flácido incapaz de sustentar a cabeça, a qual apoia-se no piso, desprendimento das penas e paralisia flácida das asas. A toxina C, produzida pelo Clostridium botulinum, é a responsável pela doença. O presente trabalho relata casos de botulismo em aves domésticas, ocorridos na região de Conquista ? MG
Botulism is a fatal paralyses caused by the Clostridium botulinum toxin. It is observed in the majority of the avian species except vultures, which are resistant. The disease appears in birds and presents the following clinical signs: flaccid neck incapable of supporting the head, which is supported on the ground, dropping feathers, and flaccid paralysis involving the wings. Toxin-C, which is produced by Clostridium botulinum, is responsible for the disease. The present work relates botulism cases in birds, occurred in the region city of Conquista, MG, Brazil
Botulismo es una enfermedad que provoca una parálisis fatal, causada por la toxina del Clostridium botulinum. El botulismo es observado en la mayoría de las especies avícolas, excepto los buitres, los cuales son resistentes. La enfermedad se manifiesta en las aves, presentado los siguientes señales clínicos: cuello flácido incapaz de sostener la cabeza, apoyándose en el suelo, caída de las plumas y parálisis flácida de las alas. La toxina C, producida por el Clostridium botulinum, es La responsable por la enfermedad. Esta investigación relata casos de botulismo en aves domésticas, ocurridos en la región de Conquista ? MG