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PMID: 2537955 Published · ppublish English Journal Article

Cloning the BamHI restriction modification system.

Nucleic acids research ·Vol. 17 ·No. 3 ·1989-02-11 ·Pages 979-97

Brooks JE, Benner JS, Heiter DF, Silber KR, Sznyter LA, Jager-Quinton T, Moran LS, Slatko BE, Wilson GG, Nwankwo DO

Abstract

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.

MeSH Terms
Amino Acid Sequence Bacillus/enzymology,genetics Bacterial Proteins/genetics,isolation & purification Base Sequence Cloning, Molecular/methods DNA Modification Methylases/genetics DNA Probes/genetics,isolation & purification DNA Transposable Elements DNA, Bacterial/isolation & purification Deoxyribonuclease BamHI/genetics,isolation & purification,physiology Escherichia coli/genetics Gene Rearrangement Molecular Sequence Data Plasmids Restriction Mapping/methods Transformation, Genetic
Chemicals
Bacterial Proteins DNA Probes DNA Transposable Elements DNA, Bacterial DNA Modification Methylases Deoxyribonuclease BamHI
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Brooks J E
New England Biolabs, Beverly, MA 01915.
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
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.3.979
Published
1989-02-11
Pages
979-97
Language
English
Region
England
NLM ID
0411011
PMCID
PMC331717
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/17.3.979 PMC Full Text View journal details
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