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Nucleic Acids Research, 1989, Vol. 17, No. 3 979-997
© 1989

MOLECULAR BIOLOGY

Cloning the BamHI restriction modification system

Joan E. Brooks, Jack S. Benner, Daniel F. Heiter, Karen R. Silber*, Laura A. Sznyter, Tina Jager-Quinton, Laurie S. Moran, Barton E. Slatko, Geoffrey G. Wilson and Donald O. Nwankwo

New England Biolabs Beverly, MA 01915, USA

Received November 4, 1988. Revised December 21, 1988. Accepted December 21, 1988.

BamHI, a Type II restriction modification system from Bacillus amyloliquefaciensH recognizes the sequence GGATCC. The methylase and endonuclease genes have been cloned into E. coli in separate steps; the clone is able to restrict unmodified phage. Although within the clone the methylase and endonuclease genes are present on the same pACYC184 vector, the system can be maintained in E. coli only with an additional copy of the methylase gene present on a separate vector.

The initial selection for BamHI methylase activity also yielded a second BamHI methylase gene which is not homologous in DNA sequence and hybridizes to different genomic restriction fragments than does the endonuclease-linked methylase gene.

Finally, the interaction of the BamHI system with the E. coli Dam and the Mcr A and B functions, have been studied and are reported here.

*Present address: Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA


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