Molecular characterization of a gene encoding a 72-kilodalton mosquito-toxic crystal protein from Bacillus thuringiensis subsp. israelensis

Donovan, William P.; Dankocsik, C C; Gilbert, Michel

doi:10.1128/jb.170.10.4732-4738.1988

Cited by 112 publications

(59 citation statements)

References 36 publications

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“…It was not possible to correlate the aggregation phenotype with the presence of a particular plasmid (data not presented). The recipient strains (AND606 and AND706) are spontaneous acrystalliferous derivatives of AND406 and lack the 75-MDa plasmid (also referred to as 112-kb plasmid [39] or 108-kb plasmid [10]) containing the toxin genes (14,21,39,45). Also, the 68-MDa plasmid which has been associated with the production of satellite inclusions (44) was absent in the recipient strains.…”

Section: Resultsmentioning

confidence: 99%

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Mobilization of small plasmids in Bacillus thuringiensis subsp. israelensis is accompanied by specific aggregation

1993

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Mobilizations of pBC16 and pAND006, containing the replicon of the Bacillus thuringiensis subsp. israelensis plasmid pTX14-3, between strains of B. thuringiensis subsp. israelensis were examined. Transconjugants appeared after a few minutes and reached a maximum frequency after approximately 2 h. Plasmid pBC16 was mobilized at a frequency approximately 200 times that of pAND006. However, pAND006 was consistently transferred, suggesting that the replicon of pTX14-3 is sufficient to sustain mobilization in B. thuringiensis subsp. israelensis. A specific protease-sensitive coaggregation between strains of B. thuringiensis subsp. israelensis was found to be unambiguously correlated with plasmid transfer. Two aggregation phenotypes, Agr+ and Agr-, were identified in this subspecies. Aggregation disappeared when the optical density of the mating mixture at 600 nm exceeded approximately 1, and it did not reappear upon dilution. Aggregation was shown to involve interactions of cells with opposite aggregation phenotypes, and evidence of a proteinaceous molecule on the surface of the Agr-that is cells involved in aggregation formation is presented. Matings and selection for the presence of two antibiotic resistance plasmids followed by identification of the host cell revealed that mobilization was unidirectional, from the Agr+ cell to the Agr-cell. The aggregation phenotype was found to be transferred with high frequency ("100%) in broth matings, and the appearance of Agr-isolates from Agr+ strains suggested that the loci involved in aggregation formation are located on a plasmid. No excreted aggregation-inducing signals were detected in the supernatant or culture filtrate of either the donor, the recipient, or the mating mixture.Conjugation, the direct transfer of genes from one bacterial cell to another, is often mediated by large conjugative plasmids. These large plasmids are transferred at high frequency, and occasionally chromosomal genes and smaller nonconjugative plasmids are cotransferred by the transfer apparatus of the conjugative plasmids. In gram-negative bacteria many small nonconjugative plasmids can be transferred (mobilized) if an additional self-transmissible plasmid is present in the host (13, 46, 47). Recent reports have described similar mobilizations of small plasmids in gram-positive bacteria (2,5,26,34,40). The first studied and best understood of the conjugative plasmids is the F plasmid, which can replicate in Escherichia coli and certain closely related enteric bacteria. The mechanism of F plasmid-mediated conjugation involves physical contact of the cells and transfer of plasmid DNA brought about by the sex pilus (23). Studies of the gram-positive bacterium Enterococcus (formerly Streptococcus) faecalis suggest a mechanism quite different from that in the better-studied gram-negative systems. Pili do not play any role in conjugation between strains of this organism, but pheromone-induced aggregation between cells has been shown to be a major characteristic (11,15).

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Section: Resultsmentioning

confidence: 99%

“…This subspecies has been reported to contain up to 10 plasmids (8,21). Two of the small plasmids, pTX14-1 and pTX14-3, have been cloned and characterized (3,6,30), a 75-MDa plasmid has been shown to encode the toxin genes (14,21,39,45), and the transfer functions have been ascribed to a 135-MDa plasmid (21) (presumedly the same as pXO16 [36]). …”

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confidence: 99%

Mobilization of small plasmids in Bacillus thuringiensis subsp. israelensis is accompanied by specific aggregation

1993

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“…Previously two groups have sequenced the cry11A gene from B. thuringiensis subsp. israelensis 26) and from B. thuringiensis subsp. morrisoni.…”

Section: Resultsmentioning

confidence: 99%

Active Form of Dipteran-Specific Insecticidal Protein Cry11A Produced byBacillus thuringiensissubsp.israelensis

Yamagiwa

Ogawa

Yasuda

et al. 2002

Bioscience, Biotechnology, and Biochemistry

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“…pHT643, which carries the p19, c r y l l A and p20 genes, has already been described (Dervyn et al, 1995 ; The second primer (5' GAAAATAACCCTTGGCTG 3') is complementary to nucleotides overlapping the StyI site in the coding sequence of cry1lA. pEG217 was used as the template (Donovan et al, 1988). The 1191 bp PCR product was digested with BamHI and StyI, purified and ligated to pHT643 digested with BamHI and StyI.…”

Section: Methodsmentioning

confidence: 99%

SpoOA represses transcription of the cry toxin genes in Bacillus thuringiensis

et al. 1997

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The DNA regions upstream from the genes encoding polypeptides of Bacillus thuringiensis subsp. israelensis larvicidal crystals (cry4A, cry4B, cry7 7A) contain sequences with similarities to the spoOA box of Bacillus subtilis (or 'OA' box) and the promoter recognized by the #-associated RNA polymerase of B. subtilis. Expression of cry-lacZ transcriptional fusions was analysed in various B. thuringiensis genetic backgrounds. The early transcription of the toxin genes was not sporulation-dependent, whereas the late-stage expression at tM was &dependent. Primer extension analysis confirmed that the c r y 4 and cry7 7-type toxin genes were weakly transcribed during the transition phase; expression analysis of a crylIA'-lacZ transcriptional fusion in B. subtilis sporulation mutants confirmed the involvement of the #-RNA polymerase. Primer extension analysis showed that in B. thuringiensis subsp. israelensis, the cry4A and cry77A gene transcription observed at the end of the growth stage was turned off at the beginning of the sporulation phase. The DNA region located upstream from the cry77A gene promoter including the putative 'OA' box was deleted. This led to a derepression of the expression of the cry77A operon. These results suggest that the cry4A, cry4B and cry77A toxin genes of B. thuringiensis subsp. israelensis are transcribed during the transition phase by the RNA polymerase associated with the # factor and are subject t o SpoOA repression.

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Molecular characterization of a gene encoding a 72-kilodalton mosquito-toxic crystal protein from Bacillus thuringiensis subsp. israelensis

Cited by 112 publications

References 36 publications

Mobilization of small plasmids in Bacillus thuringiensis subsp. israelensis is accompanied by specific aggregation

Mobilization of small plasmids in Bacillus thuringiensis subsp. israelensis is accompanied by specific aggregation

Active Form of Dipteran-Specific Insecticidal Protein Cry11A Produced byBacillus thuringiensissubsp.israelensis

SpoOA represses transcription of the cry toxin genes in Bacillus thuringiensis

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