Cloning the BamHI restriction modification system

• Published in: Nucleic acids research Vol. 17; no. 3; pp. 979 - 997
• Main Authors: BROOKS, J. E, BENNER, J. S, HEITER, D. F, SILBER, K. R, SZNYTER, L. A, JAGER-QUINTON, T, MORAN, L. S, SLATKO, B. E, WILSON, G. G, NWANKWO, D. O
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 11-02-1989
• Subjects: Amino Acid Sequence; Bacillus; Bacillus - enzymology; Bacillus - genetics; Bacillus amyloliquefaciens; Bacterial Proteins - genetics; Bacterial Proteins - isolation & purification; Base Sequence; Biological and medical sciences; Cloning, Molecular - methods; Deoxyribonuclease BamHI - genetics; Deoxyribonuclease BamHI - isolation & purification; Deoxyribonuclease BamHI - physiology; DNA Modification Methylases - genetics; DNA Probes - genetics; DNA Probes - isolation & purification; DNA Transposable Elements; DNA, Bacterial - isolation & purification; Escherichia coli; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Gene Rearrangement; Genes. Genome; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Plasmids; Restriction Mapping - methods; Transformation, Genetic; Endodeoxyribonuclease BamHI; Gene; Nucleotide sequence; Escherichia coli; Restriction map; Bacteria; Molecular cloning; Southern blotting; Escherichieae; Enterobacteriaceae
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/17.3.979

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.