Expression of Bacillus thuringiensis delta-endotoxin genes during vegetative growth

Mettus, A M; Macaluso, A

doi:10.1128/aem.56.4.1128-1134.1990

Cited by 31 publications

(11 citation statements)

References 24 publications

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“…Symbols: Lii, vector sequences derived from pBR322; , vector sequences derived from pNN101. megaterium and B. thuringiensis (29). Insertions within the tet structural gene (as in pEG93) are probably also expressed during vegetative growth and may serve as the basis for the increased accumulation of toxin in VT1660(pEG93).…”

Section: Resultsmentioning

confidence: 99%

Insecticidal toxins from Bacillus thuringiensis subsp. kenyae: gene cloning and characterization and comparison with B. thuringiensis subsp. kurstaki CryIA(c) toxins

Tersch

Robbins

Jany

et al. 1991

Appl Environ Microbiol

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Genes encoding insecticidal crystal proteins were cloned from three strains of Bacillus thuringiensis subsp. kenyae and two strains of B. thuringiensis subsp. kurstaki. Characterization of the B. thuringiensis subsp. kenyae toxin genes showed that they are most closely related to cryIA(c) from B. thuringiensis subsp. kurstaki. The cloned genes were introduced into Bacillus host strains, and the spectra of insecticidal activities of each Cry protein were determined for six pest lepidopteran insects. CryIA(c) proteins from B. thuringiensis subsp. kenyae are as active as CryIA(c) proteins from B. thuringiensis subsp. kurstaki against Trichoplusia ni, Lymantria dispar, Heliothis zea, and H. virescens but are significantly less active against Plutella xylostella and, in some cases, Ostrinia nubilalis. The sequence of a crylA(c) gene from B. thuringiensis subsp. kenyae was determined (GenBank M35524) and compared with that of crylA(c) from B. thuringiensis subsp. kurstaki. The two genes are more than 99% identical and show seven amino acid differences among the predicted sequences of 1,177 amino acids.

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Section: Resultsmentioning

confidence: 99%

Insecticidal toxins from Bacillus thuringiensis subsp. kenyae: gene cloning and characterization and comparison with B. thuringiensis subsp. kurstaki CryIA(c) toxins

Tersch

Robbins

Jany

et al. 1991

Appl Environ Microbiol

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“…Macaluso and Mettus (14) used the promoter of the tetracycline resistance gene (tet) to express genes cryL4(c) and cryILA during vegetative growth, but the expression level was relatively low. By using their system, cry gene expression was extended from the vegetative stage to the sporulation stage.…”

Section: Discussionmentioning

confidence: 99%

“…Several reports have described the used of vegetative promoters for the expression of cry genes in Escherichia coli (14), B. stearothermophilus (16), and B. thuringiensis (12,14) strains. However, these systems exhibited low-level expression of the newly introduced cry gene and/or interference with expression of the resident cry gene(s).…”

mentioning

confidence: 99%

Expression of the crystal protein gene under the control of the alpha-amylase promoter in Bacillus thuringiensis strains

Chak¹,

Tseng²,

Yamamoto³

1994

Appl Environ Microbiol

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The expression of an insecticidal crystal protein gene of Bacillus thuringiensis under the control of the a-amylase gene promoter was investigated. The crylC gene, which encodes a protein known to have a unique activity against Spodoptera (armyworm) species, was used in this investigation. The cryIC gene was placed, along with the a-amylase promoter from B. subtilis, in a B. thuringiensis-derived cloning vector, generating a pair of recombinant plasmids, pSB744 and pSB745. The cloning vector that contains the minimal replicon of B. thuringiensis subsp. kurstaki HD73 is stably maintained in a variety of B. thuringiensis strains, as previously reported by Gamel and Piot (Gene 120:17-26, 1992). The present study confirmed that the recombinant plasmids are also stably maintained in B. thuringiensis subsp. kurstaki Cry-B and HD73 growing in media without selection pressure for at least 48 h. The cryIC gene on the recombinant plasmids were notably

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“…E. coli transformations were performed using frozen competent cells as described by the manufacturer (GIBCO BRL or Stratagene Cloning Systems). B. thuringiensis was transformed as described by Mettus and Macaluso (23). Plasmid isolations from E. coli and B. thuningiensis were by the alkaline lysis procedure described by Maniatis et al (21).…”

Section: Methodsmentioning

confidence: 99%

Overexpression of Bacillus thuringiensis HknA, a histidine protein kinase homology, bypasses early Spo mutations that result in CryIIIA overproduction

1994

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The Bacillus thuringiensis CryIIIA insecticidal crystal protein (ICP) is a vegetatively expressed protein that is toxic to coleopteran insect larvae. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the asporogenous B. thuringiensis subsp. morrisoni strain EG1351, which harbors the native cryIIIA-encoding 88-MDa plasmid, showed a 2.5-fold overproduction of the CryIIIA protein compared with that of an isogenic wild-type strain. Further studies showed that neither CryIIIA protein synthesis nor CryIIIA protein processing was affected in strain EG1351 during vegetative growth. In an attempt to characterize the EG1351 mutation by complementation of function, the hknA gene was identified and cloned from a B. thuringiensis cosmid library. Primer extension analysis of hknA mRNA in wild-type B. thuringiensis demonstrated that the hknA gene is transcribed during vegetative growth from a sigma A-like promoter. Multiple copies of either the hknA gene or the Bacillus subtilis kinA (spoIIJ) gene were shown to bypass the sporulation defect in strain EG1351 as well as a spo0F mutation in B. thuringiensis EG1634. Additional studies showed that the hknA gene was not defective in strain EG1351. The results of this study suggest that hknA encodes a novel histidine protein kinase involved in B. thuringiensis sporulation. We also propose that the CryIIIA-overproducing phenotype of strain EG1351 is most likely due to a defect in the phosphorylation of Spo0A and confirm that CryIIIA production is not dependent on sporulation.

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Expression of Bacillus thuringiensis delta-endotoxin genes during vegetative growth

Cited by 31 publications

References 24 publications

Insecticidal toxins from Bacillus thuringiensis subsp. kenyae: gene cloning and characterization and comparison with B. thuringiensis subsp. kurstaki CryIA(c) toxins

Insecticidal toxins from Bacillus thuringiensis subsp. kenyae: gene cloning and characterization and comparison with B. thuringiensis subsp. kurstaki CryIA(c) toxins

Expression of the crystal protein gene under the control of the alpha-amylase promoter in Bacillus thuringiensis strains

Overexpression of Bacillus thuringiensis HknA, a histidine protein kinase homology, bypasses early Spo mutations that result in CryIIIA overproduction

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