Screening for novel cry genes by hybridization

Lett Appl Microbiol. 2001 Sep;33(3):241-5. doi: 10.1046/j.1472-765x.2001.00982.x.

Abstract

Aims: To develop an efficient and sensitive method to facilitate the search for novel Cry toxins.

Methods and results: The method uses a cocktail of cry gene sequences as a hybridization probe to screen Bacillus thuringiensis (Bt) strains and gene libraries prepared from them. Under the hybridization and washing conditions used, cross-hybridization between genes from different cry families was not observed. Probes containing either partial or complete cry gene sequences produced similar patterns when hybridized to genomic DNA of several Bt strains although the pattern produced by the probe composed of entire gene coding regions was somewhat more complex.

Conclusion: As a tool for gene library screening, hybridization with a mixture of cry gene sequences is an efficient means of detecting clones containing a diverse range of cry genes in a single step.

Significance and impact of the study: This technique greatly improves the ease and efficiency of novel toxin gene discovery compared to previous methods.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus thuringiensis / genetics*
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / genetics*
  • Bacterial Toxins / genetics
  • DNA Probes
  • Endotoxins / genetics*
  • Genes, Bacterial*
  • Genomic Library
  • Hemolysin Proteins
  • Nucleic Acid Hybridization*
  • Polymerase Chain Reaction
  • Sensitivity and Specificity

Substances

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