Summary
We took advantage of the recently discovered high frequency transfer of plasmids between strains of B. thuringiensis to obtain the heterospecific mating between a Bacillus subtilis strain, which contained the plasmid crystal gene from strain berliner 1715, and different B. thuringiensis strains. The plasmid-coded crystal gene was inserted in the plasmid pBT 42-1 and introduced into an acrystalliferous B. thuringiensis strain where it promoted the synthesis of the crystal protein. The plasmid was maintained stably and allowed the synthesis of a toxic parasporal body. The plasmid pBT 42-1 was also introduced into a wild-type strain israelensis and the transcipient strains produced both types of δ endotoxin, which are active on both lepidopteran and dipteran larvae. The transfers were also performed using the cloned crystal gene of chromosomal origin, but in this case, the gene was integrated into the chromosomal DNA of the transcipient strains and did not seem to be expressed.
Similar content being viewed by others
References
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl Acids Res 7:1513–1523
Bulla LA, Kramer KT, Davidson LI (1977) Characterization of the entomocidal parasporal crystal of B. thuringiensis. J Bacteriol 130:375–383
Charles JF, de Barjac H (1981) Sporulation et cristallogénèse de B. thuringiensis var. israelensis en microscopic électronique. Ann Inst Pasteur 133A:425–438
de Barjac H (1978) Une nouvelle variété de Bacillus thuringiensis très toxique pour les moustiques: B. thuringiensis var. israelensis, sérotype H-14. CR Acad Sci Paris (série D) 286:787–800
Dulmage HT (1979) In: Hoy MA, McKelvey JJ Jr (eds) Genetics in relation to insect management. Rockefeller Foundation, New York, p 116
Gonzalez JM, Dulmage HT, Carlton BC (1981) Correlation between specific plasmids and δ endotoxin production in B. thuringiensis. Plasmid 5:351–365
Gonzalez JM, Brown BJ, Carlton BC (1982) Transfer of B. thuringiensis plasmid coding for δ endotoxin among strains of B. thuringiensis and B. cereus. Proc Natl Acad Sci USA 79:6951–6955
Held GA, Bulla LA, Ferrari E, Hoch J, Aronson AI (1982) Cloning and localization of the lepidopteran protoxin gene of B. thuringiensis subsp. kurstaki. Proc Natl Acad Sci USA 79:6065–6069
Klier A, Fargette F, Ribier J, Rapoport G (1982) Cloning and expression of the crystal protein genes from B. thuringiensis strain berliner 1715. EMBO J 1:791–799
Klier A, Parsot C, Rapoport G (1983) In vitro transcription of the cloned chromosomal crystal gene from B. thuringiensis. Nucl Acids Res (In press)
Lereclus D, Lecadet M-M, Ribier J, Dedonder R (1981) Molecular relationships among plasmids of B. thuringiensis: conserved sequences through 11 crystalliferous strains. Mol Gen Genet 186:391–398
Rapoport G, Klier A, Billault A, Fargette F, Dedonder R (1979) Construction of a colony bank of E. coli containing hybrid plasmids representative of the B. subtilis 168 genome. Mol Gen Genet 176:239–245
Ribier J, Lecadet M-M (1973) Etude ultrastructurale et cinétique de la sporulation de Bacillus thuringiensis var. berliner 1715. Remarques sur la formation de l'inclusion parasporale. Ann Microbiol (Institut Pasteur) 124A:311–344
Rigby DWJ, Dieckman M, Rhodes C, Berg P (1977) Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Schnepf AE, Whiteley HR (1981) Cloning and expression of the B. thuringiensis crystal protein gene in E. coli. Proc Natl Acad Sci USA 78:2893–2897
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Author information
Authors and Affiliations
Additional information
Communicated by A. Böck
Rights and permissions
About this article
Cite this article
Klier, A., Bourgouin, C. & Rapoport, G. Mating between Bacillus subtilis and Bacillus thuringiensis and transfer of cloned crystal genes. Mol Gen Genet 191, 257–262 (1983). https://doi.org/10.1007/BF00334823
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00334823