Accumulation of the insecticidal crystal protein of Bacillus thuringiensis subsp. kurstaki in post-exponential-phase Bacillus subtilis.
A gene from Bacillus thuringiensis subsp. kurstaki that codes for a Lepidoptera-specific insecticidal toxin (delta-endotoxin) was engineered for expression in Bacillus subtilis. A low-copy-number plasmid vector that replicates in Escherichia coli and B. subtilis was constructed to transform B. subtilis with gene fusions first isolated and characterized in E. coli. Naturally occurring promoter sequences from B. subtilis (43, veg, ctc, and spoVG) were inserted upstream from the plasmid-borne structural gene. In the most prolific case, when the sporulation-specific spoVG promoter was fused to the heterologous toxin gene, the toxin product accumulated during postexponential growth to greater than 25% of the total cell protein. However, the resulting specific activity of the insecticidal toxin product was not commensurate with the abundance of the protein.
Crissman
J W
JW
Syntro Corporation, San Diego, California 92121.
Causey
S C
SC
Thorne
L
L
Pollock
T J
TJ
D000818:Animals; D001412:Bacillus subtilis / Q000235:genetics / Q000254:growth & development*; D001413:Bacillus thuringiensis / Q000235:genetics / Q000378:metabolism*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity; D001427:Bacterial Toxins; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004731:Endotoxins; D005822:Genetic Vectors; D006460:Hemolysin Proteins; D007300:Insect Control; D007814:Larva / Q000187:drug effects / Q000382:microbiology; D007915:Lepidoptera / Q000187:drug effects / Q000382:microbiology*; D010957:Plasmids; D011401:Promoter Regions, Genetic
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis