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. 1984 Nov;3(11):2561–2567. doi: 10.1002/j.1460-2075.1984.tb02174.x

A transposon-like structure related to the delta-endotoxin gene of Bacillus thuringiensis.

D Lereclus, J Ribier, A Klier, G Menou, M M Lecadet
PMCID: PMC557730  PMID: 6096131

Abstract

A DNA segment (Th-sequence) has been found in several strains of Bacillus thuringiensis. This Th-sequence [3 megadaltons (Md)] induces adjacent deletions when it is located in the pAM beta 1 plasmid derived from Streptococcus faecalis. Electron microscopic examination of reannealed single strands of one plasmid (pMT9) carrying such a deletion revealed that the Th-sequence corresponds to a single-stranded loop (2.8 Md) bounded by a short double-stranded stem (less than 0.2 Md). Southern blotting experiments established that in B. thuringiensis the Th-sequence was generally located on the large plasmid which also harbours the gene coding for the delta-endotoxin (crystal protein). Hybridization and heteroduplex analysis of the extrachromosomal DNA from the berliner 1715 strain demonstrated that the crystal gene and the Th-sequence are located in close vicinity on a 42-Md plasmid and that they are separated by a 1.3-Md DNA segment. This DNA segment is repeated in inverted orientation, once immediately adjacent to the Th-sequence and once 1.8 Md beyond the crystal gene. A model for the organization of these DNA sequences inside a transposon-like structure is proposed.

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Selected References

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  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Held G. A., Bulla L. A., Jr, Ferrari E., Hoch J., Aronson A. I., Minnich S. A. Cloning and localization of the lepidopteran protoxin gene of Bacillus thuringiensis subsp. kurstaki. Proc Natl Acad Sci U S A. 1982 Oct;79(19):6065–6069. doi: 10.1073/pnas.79.19.6065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Khan S. A., Novick R. P. Terminal nucleotide sequences of Tn551, a transposon specifying erythromycin resistance in Staphylococcus aureus: homology with Tn3. Plasmid. 1980 Sep;4(2):148–154. doi: 10.1016/0147-619x(80)90004-9. [DOI] [PubMed] [Google Scholar]
  4. Klier A. F., Lecadet M. M., Dedonder R. Sequential modifications of DNA-dependent RNA polymerase during sporogenesis in Bacillus thuringiensis. Eur J Biochem. 1973 Jul 16;36(2):317–327. doi: 10.1111/j.1432-1033.1973.tb02916.x. [DOI] [PubMed] [Google Scholar]
  5. Klier A., Fargette F., Ribier J., Rapoport G. Cloning and expression of the crystal protein genes from Bacillus thuringiensis strain berliner 1715. EMBO J. 1982;1(7):791–799. doi: 10.1002/j.1460-2075.1982.tb01249.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Klier A., Parsot C., Rapoport G. In vitro transcription of the cloned chromosomal crystal gene from Bacillus thuringiensis. Nucleic Acids Res. 1983 Jun 25;11(12):3973–3987. doi: 10.1093/nar/11.12.3973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kronstad J. W., Schnepf H. E., Whiteley H. R. Diversity of locations for Bacillus thuringiensis crystal protein genes. J Bacteriol. 1983 Apr;154(1):419–428. doi: 10.1128/jb.154.1.419-428.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Leblanc D. J., Lee L. N. Physical and genetic analyses of streptococcal plasmid pAM beta 1 and cloning of its replication region. J Bacteriol. 1984 Feb;157(2):445–453. doi: 10.1128/jb.157.2.445-453.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lereclus D., Lecadet M. M., Ribier J., Dedonder R. Molecular relationships among plasmids of Bacillus thuringiensis: conserved sequences through 11 crystalliferous strains. Mol Gen Genet. 1982;186(3):391–398. doi: 10.1007/BF00729459. [DOI] [PubMed] [Google Scholar]
  10. Lereclus D., Menou G., Lecadet M. M. Isolation of a DNA sequence related to several plasmids from Bacillus thuringiensis after a mating involving the Streptococcus faecalis plasmid pAM beta 1. Mol Gen Genet. 1983;191(2):307–313. doi: 10.1007/BF00334831. [DOI] [PubMed] [Google Scholar]
  11. Niaudet B., Ehrlich S. D. In vitro genetic labeling of Bacillus subtilis cryptic plasmid pHV400. Plasmid. 1979 Jan;2(1):48–58. doi: 10.1016/0147-619x(79)90005-2. [DOI] [PubMed] [Google Scholar]
  12. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  13. Schnepf H. E., Whiteley H. R. Cloning and expression of the Bacillus thuringiensis crystal protein gene in Escherichia coli. Proc Natl Acad Sci U S A. 1981 May;78(5):2893–2897. doi: 10.1073/pnas.78.5.2893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Wong H. C., Schnepf H. E., Whiteley H. R. Transcriptional and translational start sites for the Bacillus thuringiensis crystal protein gene. J Biol Chem. 1983 Feb 10;258(3):1960–1967. [PubMed] [Google Scholar]
  15. Yamamoto T., Yokota T. Escherichia coli heat-labile enterotoxin genes are flanked by repeated deoxyribonucleic acid sequences. J Bacteriol. 1981 Feb;145(2):850–860. doi: 10.1128/jb.145.2.850-860.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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