Summary
The effects of the restriction system of Bacillus subtilis strain M on plasmid transformation were studied. Plasmid pHV1401 DNA prepared from B. subtilis transformed the restriction-proficient M strain 100 times more efficiently than the DNA prepared from Escherichia coli, while the two DNA preparations transformed restriction-deficient derivatives of that strain with similar efficiencies. This indicates that transformation with pHV1401 is sensitive to the M restriction system. pHV1401 contains three CTCGAG (XhoI sites). Successive removal of these abolished the effect of restriction. This indicates that the XhoI sites are the targets for the M restriction system.
This is a preview of subscription content, log in via an institution to check access.
Abbreviations
- Apr :
-
resistance to ampicillin
- Cmr :
-
resistance to chloramphenicol
- R/M:
-
restriction and modification
- Tcr :
-
resistance to tetracycline
References
Auerswald EA, Ludwig G, Schaller H (1980) Structural analysis of Tn5. Cold Spring Harbor Symp Quant Biol 45:107–113
Beck E, Ludwig G, Auerswald EA, Reiss B, Schaller H (1982) Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene 19:327–336
Bron S, Luxen E, Venema G, Trautner TA (1980a) Restriction and modification in B. subtilis. Effects on transformation and transfection with native and single-stranded DNA. Mol Gen Genet 179:103–110
Bron S, Luxen E, Trautner TA (1980b) Restriction and modification in B. subtilis. The role of homology between donor and recipient DNA in transformation and transfection. Mol Gen Genet 179:111–117
Canosi U, Lüder G, Trautner TA, Bron S (1981) Restriction and modification in B. subtilis: effects on plasmid transformation. In: Polsinelli M (ed) Proceedings of the 5th European Meeting on transformation and transfection. Cotswood Press Ltd, Oxford, pp 179–187
Guha S (1985) Determination of DNA sequences containing methylcytosine in Bacillus subtilis Marburg. J Bacteriol 163:573–579
Haima P, Bron S, Venema G (1987) The effect of restriction on shotgun cloning and plasmid stability in Bacillus subtilis Marburg. Mol Gen Genet 209:335–342
Ikawa S, Shibata T, Ando T, Saito H (1980) Genetic studies on site-specific endodeoxyribonucleases in Bacillus subtilis: multiple modification and restriction systems in transformants of Bacillus subtilis 168. Mol Gen Genet 177:359–368
Jannière L, Ehrlich SD (1987) Recombination between short repeated sequences is more frequent in plasmids than in the chromosome of Bacillus subtilis. Mol Gen Genet 210:116–121
Jentsch S (1983) Restriction and modification in Bacillus subtilis: sequence specificities of restriction/modification systems BsuM, BsuE and BsuF. J Bacteriol 156:800–808
Mazodier P, Cossart P, Giraud E, Gasser F (1985) Completion of the nucleotide sequence of the central region of Tn5 confirms the presence of three resistance genes. Nucleic Acids Res 13:195–205
Niaudet B, Ehrlich SD (1979) In vitro genetic labeling of Bacillus subtilis cryptic plasmid pHV400. Plasmid 2:48–58
Ostroff GR, Pène JJ (1983) Molecular cloning with bifunctional plasmid vectors in Bacillus subtilis III. Isolation of a spontaneous mutant of B. subtilis with enhanced transformability for Escherichia coli-propagated chimeric plasmid DNA. J Bacteriol 156:934–936
Ostroff GR, Pène JJ (1984a) Molecular cloning with bifunctional plasmid vectors in Bacillus subtilis. II. Transfer of sequences propagated in Escherichia coli to B. subtilis. Mol Gen Genet 193:306–311
Ostroff GR, Pène JJ (1984b) Parameters affecting the stability of recombinant bifunctional plasmid transfer from Escherichia coli to Bacillus subtilis. In: Ganesan AT, Hoch JA (eds) Genetics and Biotechnology of Bacilli. Academic Press, New York, pp 285–295
Primrose SB, Ehrlich SD (1981) Isolation of plasmid deletion mutants and study of their instability. Plasmid 6:193–201
Shibata T, Ando T (1974) Host controlled modification and restriction in Bacillus subtilis. Mol Gen Genet 131:275–280
Tanaka T (1979) Restriction of plasmid-mediated transformation in Bacillus subtilis. Mol Gen Genet 175:235–237
Trautner TA, Pawlek B, Bron S, Anagnostopoulos C (1974) Restriction and modification in B. subtilis. Biological aspects. Mol Gen Genet 131:181–191
Uozumi T, Hoshino T, Miwa K, Horinouchi S, Beppu T, Arima K (1977) Restriction and modification in Bacillus species. Genetic transformation of bacteria with DNA from different species, part I. Mol Gen Genet 152:65–69
Author information
Authors and Affiliations
Additional information
Communicated by W. Gajewski
Rights and permissions
About this article
Cite this article
Bron, S., Jannière, L. & Ehrlich, S.D. Restriction and modification in Bacillus subtilis Marburg 168: Target sites and effects on plasmid transformation. Mol Gen Genet 211, 186–189 (1988). https://doi.org/10.1007/BF00338412
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00338412