Toxin A of Clastr~dium d~~cile has been purified and monospecific antiserum produced. A reliable procedure for isolation and restriction of C. dl~ci~e chromosomal DNA was developed which allowed for the construction of a genomic library in d gtll. Approx. 35 000 plaques were screened using anti-toxin A which resulted in the identification of one stable positive clone, d cd19. Verification of the immunological identity of the isolated toxin A gene fragment in A cdl9 was determined by affinity purifying toxin A antibodies specific for 1 cdl9 gene product, and using these selected antibodies to probe a Western blot of purified toxin A. The insert in 1 cdl9 was demonstrated to be a 0.3 kb fragment by restriction digestion, and by hybridization of the clone to a chromosomal digest of C. d@cile. The peptide coded for by the toxin A gene fragment in 1 cdl9 was not cytotoxic for 3T3 mammalian tissue culture cells.
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