Molecular cloning of the delta-endotoxin gene of Bacillus thuringiensis var. israelensis

Gene. 1985;33(2):151-8. doi: 10.1016/0378-1119(85)90089-7.

Abstract

A transformant of Bacillus megaterium, VB131, was isolated which carries a 6.3-kb XbaI segment of the crystal toxin gene of Bacillus thuringiensis var. israelensis (BTI) cloned in a vector plasmid pBC16 to yield pVB131. The chimeric plasmid DNA from VB131 was introduced into a transformable Bacillus subtilis strain by competence transformation. Both the B. megaterium VB131 strain and the B. subtilis strain harboring the chimeric plasmid produced irregular, parasporal, phase-refractile, crystalline inclusions (Cry+) during sporulation. The sporulated cells as well as the isolated crystal inclusions of the pVB131-containing B. megaterium and B. subtilis strains were highly toxic to the larvae of Aedes aegypti. Also, the solubilized crystal protein preparation from VB131[pVB131] showed clear immuno cross-reaction with antiserum to the BTI crystal toxin. 32P-labeled pVB131 plasmid DNA showed specific hybridization with a 112-kb plasmid DNA of Cry+ strains of BTI, and no hybridization with other plasmid or chromosomal DNA of either Cry+ or Cry- variants. These results are in agreement with our previous findings (González and Carlton, 1984) that the 112-kb plasmid of BTI is associated with the production of the crystal toxin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Bacterial / genetics
  • Bacillus megaterium / genetics
  • Bacillus thuringiensis / genetics*
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / genetics
  • Bacterial Proteins / immunology
  • Bacterial Toxins*
  • Biological Assay
  • Cloning, Molecular
  • DNA, Recombinant
  • Endotoxins / genetics*
  • Genes
  • Genes, Bacterial
  • Hemolysin Proteins

Substances

  • Antigens, Bacterial
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins
  • Bacterial Toxins
  • DNA, Recombinant
  • Endotoxins
  • Hemolysin Proteins
  • insecticidal crystal protein, Bacillus Thuringiensis