Characterization and cloning of MwoI (GCN7GC), a new type-II restriction-modification system from Methanobacterium wolfei.
R.MwoI, a type-II restriction enzyme with the new specificity 5'-GCN7GC-3', was found in extracts of the thermophilic archaebacterium, Methanobacterium wolfei. R.MwoI cleaves duplex DNA producing fragments with 3-nt, 3'-terminal extensions, thus: GCN5/N2GC. The genes coding for the MwoI restriction and modification enzymes were cloned into Escherichia coli on the plasmid vector pBR322. The clones synthesize a low level of R.MwoI endonuclease. The plasmids display incomplete MwoI-specific modification, suggesting that the clones synthesize a low level of the M.MwoI methyltransferase, too.
Lunnen
K D
KD
New England Biolabs, Inc., Beverly, MA 01915.
Morgan
R D
RD
Timan
C J
CJ
Krzycki
J A
JA
Reeve
J N
JN
Wilson
G G
GG
D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000302:isolation & purification; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D019605:Euryarchaeota / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D005798:Genes, Bacterial; D010957:Plasmids; D013379:Substrate Specificity
D004269:DNA, Bacterial; C060413:endodeoxyribonuclease MwoI; D015252:Deoxyribonucleases, Type II Site-Specific