Genetic Engineering of Clostridium Difficile Toxin A Vaccine

摘要

Recombinant DNA technology allows for an approach to subunit vaccine productionthat should provide advantages over existing techniques. The major long range objective of this project is directed toward the improvement of vaccine biotechnology in the area of recombinant DNA studies using Clostridium difficile toxin A as the model. This was proposed to be accomplished by using a unique series of recombinant DNA techniques to map epitopes. Recombinant DNA technology allows for an approach to subunit vaccine production that should provide advantages over existing techniques. Improvement of vaccine biotechnology in the area of recombinant DNA studies using Clostridium difficile toxin A as the model, is the major long range objective of this project. A genomic library of C. difficile chromosomal DNA was screened using anti-toxin A which resulted in the identification of one stable positive clone. The insert was demonstrated to be a 0.3 kb fragment by restriction digestion, and by hybridization of the clone to a chromosomal digest of C. difficile. Verification of the immunological identify of the isolated toxin A gene fragment was determined by affinity purifying toxin A antibodies and using these selected antibodies to probe a Western blot of purified toxin A.