Cloning the BamHI restriction modification system.
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.
Brooks
J E
JE
New England Biolabs, Beverly, MA 01915.
Benner
J S
JS
Heiter
D F
DF
Silber
K R
KR
Sznyter
L A
LA
Jager-Quinton
T
T
Moran
L S
LS
Slatko
B E
BE
Wilson
G G
GG
Nwankwo
D O
DO
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D015254:DNA Modification Methylases / Q000235:genetics; D015342:DNA Probes / Q000235:genetics / Q000302:isolation & purification; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000302:isolation & purification; D015245:Deoxyribonuclease BamHI / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics; D015321:Gene Rearrangement; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping / Q000379:methods; D014170:Transformation, Genetic
D001426:Bacterial Proteins; D015342:DNA Probes; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D015254:DNA Modification Methylases; D015245:Deoxyribonuclease BamHI