Screening for novel cry genes by hybridization.
OBJECTIVE
To develop an efficient and sensitive method to facilitate the search for novel Cry toxins.


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.


CONCLUSIONS
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.


CONCLUSIONS
This technique greatly improves the ease and efficiency of novel toxin gene discovery compared to previous methods.
Beard
C E
CE
CSIRO Division of Entomology, GPO Box 1700, Canberra, A.C.T., 2614, Australia.
Ranasinghe
C
C
Akhurst
R J
RJ
D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics; D015342:DNA Probes; D004731:Endotoxins / Q000235:genetics*; D005798:Genes, Bacterial; D015698:Genomic Library; D006460:Hemolysin Proteins; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D015342:DNA Probes; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis