Summary
We transformed B. subtilis 168 with DNA from B. subtilis IAM1231, IAM1192 and ATCC6633. When we examined the restriction activities of the transformants in vivo and in vitro using phage ϕ105C we found the following: (1) Cells of either IAM1231 or IAM1192 have two modification and restriction systems (Bsu1231(1)-system and Bsu1231(II)-system in IAM1231, and Bsu1192(I)-system and Bsu1192(II)-systems in IAM1192), and cells of ATCC6633 have only one system (Bsu6633-system). (2) The restriction enzymes of all of these five systems are site-specific endonucleases. (3) The nucleotide sequence specificities of the enzymes involved in Bsu1231(I)-system, Bsu1192(I)-system and Bsu6633-system are the same; and those of Bsu1231(II)-system and Bsu1192(II)-system are the same. The sequence specificities of these two groups are different from each other and also different from those of the Bsu168-system of B. subtilis 168, the BsuR-system of B. subtilis R and the Bsu1247(I)-and Bsu1247(II)-systems which are systems of B. subtilis IAM1247. (4) Transformants possessing four different modification and restriction systems (Bsu1231(I)-, Bsu1247(I)-, BsuR- and Bsu168-systems) were constructed. (5) Transformation of two derivatives of 168 that were m +R r +R by DNA from IAM1231 produced 16 transformants that had the Bsu1231(II) restriction system, but had lost the BsuR system. Transformation of a derivative of 168 that was m +1247(II) r +1247(II) by DNA from m +1231(II) r +1231(II) -or m +R r +R -derivative of 168 produced about 100 each of transformants that had the Bsu1231(II)-restriction system or the BsuR-restriction system. But all these transformants lost the Bsu1247(II)-system.
Similar content being viewed by others
References
Arber, W., Morse, M.L.: Host specificity of DNA produced by Escherichia coli. VI. Effects on Bacterial conjugation. Genetics 51, 137–148 (1965)
Bannister, D., Grover, S.W.: The isolation and properties of two different host specificities associated with drug resistance factors. J. Gen. Microbiol. 61, 63–71 (1970)
Betlach, M., Hershfield, V., Chow, L., Brown, W., Goodman, H.M., Boyer, H.W.: A restriction endonuclease analysis of the bacterial plasmid controlling the EcoRI restriction and modification of DNA. Fed. Proc. 35, 2037–2043 (1976)
Bron, S., Murray, K.: Restriction and modification in B. subtilis. Nucleotides sequence recognized by restriction endonuclease R. BsuR from strain R. Mol. Gen. Genet. 143, 25–33 (1975)
Bron, S., Murray, K., Trautner, T.A.: Restriction and modification in B. subtilis. Purification and general properties of a restriction endonuclease from strain R. Mol. Gen. Genet. 143, 13–23 (1975)
Chang, S., Cohen, S.N.: In vivo site-specific genetic recombination promoted by the EcoRI restriction endonuclease. Proc. Natl. Acad. Sci. U.S.A. 74, 4811–4815 (1977)
Ikawa, S., Shibata, T., Ando, T., Saito, H.: Host controlled modification and restriction in Bacillus subtilis. Bsu168-system and BsuR-system in B. subtilis 168. Mol. Gen. Genet. 170, 123–127 (1979)
Lacks, S., Greenberg, B.: A Deoxyribonuclease of Diplococcus pneumoniae specific for methylated DNA. J. Biol. Chem. 250, 4060–4066 (1975)
Lacks, S., Greenberg, B.: Complementary specificity of restriction endonucleases of Diplococcus pneumoniae with respect to DNA methylation. J. Mol. Biol. 114, 153–168 (1977)
Roberts, R.J.: Restriction endonucleases. Crit. Rev. Biochem. 4, 123–164 (1976)
Saito, H., Shibata, T., Ando, T.: Mapping of genes determining nonpermissiveness and host-specific restriction to bacteriophages in Bacillus subtilis Marburg. Mol. Gen. Genet. 170, 117–122 (1979)
Shibata, T., Ando, T.: Host controlled modification and restriction in Bacillus subtilis. Mol. Gen. Genet. 131, 275–280 (1974)
Shibata, T., Ando, T.: In vitro modification and restriction of phage ϕ105C DNA with Bacillus subtilis N cell-free extract. Mol. Gen. Genet. 138, 269–279 (1975)
Shibata, T., Ando, T.: The restriction endonucleases in Bacillus amyloliquefaciens N strain. Substrate specificities. Biochem. Biophys. Acta 442, 184–196 (1976)
Shibata, T., Ikawa, S., Ando, T., Saito, H.: Introduction of host controlled modification and restriction systems of B. subtilis IAM1247 into B. subtilis 168. J. Bacteriol. 139, 308–310 (1979)
Shibata, T., Ikawa, S., Kim, C., Ando, T.: Site-specific deoxyribonucleases in Bacillus subtilis and other Bacillus strains. J. Bacteriol. 128, 473–476 (1976)
Trautner, T.A., Pawlek, B., Bron, S., Anagnostopoulos, C.: Restriction and modification in B. subtilis. Biological aspect. Mol. Gen. Genet. 131, 181–191 (1974)
Uozumi, T., Hoshino, T., Miwa, K., Horinouchi, S., Beppu, T., Arima, K.: Restriction and modification in Bacillus species. Genetic transformation of bacteria with DNA from different species. Part I. Mol. Gen. Genet. 152, 65–69 (1977)
Vovis, G.F., Lacks, S.: Complementary action of restriction enzymes Endo R. DpnI and Endo R. DpnII on bacteriophage fl DNA. J. Mol. Biol. 115, 525–538 (1977)
Wilson, G.A., Young, F.E.: Isolation of a sequence-specific endonuclease (BamI) from Bacillus amyloliquefaciens H. J. Mol. Biol. 97, 123–125 (1975)
Yoshimori, R., Roulland-Dussoix, D., Boyer, H.W.: R factor-controlled restriction and modification of deoxyribonucleic acid. Restriction mutants. J. Bacteriol. 112, 1275–1279 (1972)
Author information
Authors and Affiliations
Additional information
Communicated by W. Arber
Rights and permissions
About this article
Cite this article
Ikawa, S., Shibata, T., Ando, T. et al. Genetic studies on site-specific endodeoxyribonucleases in Bacillus subtilis: Multiple modification and restriction systems in transformants of Bacillus subtilis 168. Molec. Gen. Genet. 177, 359–368 (1980). https://doi.org/10.1007/BF00271474
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00271474