1887

Abstract

, a human pathogen, produces two very large protein toxins, A and B (250–600 kDa), which resist dissociation into subunits. To clone the toxin A gene, a genomic library of 3–8 kb chromosomal DNA fragments of strain VPI 10463 established in pUC12 was screened with a rabbit polyclonal toxin A antiserum. Thirty-five clones were isolated which carried 2·5–7·0 kb inserts representing a 10 kb region of the genome. All the inserts were oriented in the same direction, suggesting that toxin A gene expression was under control of the promoter of the pUC12 vector. Western blot experiments revealed the presence of low amounts of fusion proteins of variable size (30–170 kDa) in strains harbouring recombinant plasmids. As deduced from subcloning experiments, the DNA sequences encoding toxin A comprised about 4 kb, corresponding to about 140 kDa of the 300–600 kDa protein. This was either due to incomplete cloning of the gene or it might indicate a subunit composition of toxin A. No additional gene(s) with homology to the cloned toxin A gene was detected.

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1989-01-01
2021-08-04
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