Site-Specific Methylation of Promega Restriction Enzymes

Site-Specific Methylation Sensitivity of Promega Restriction
Enzymes
Site-Specific Methylation Sensitivity of Promega Restriction Enzymes.
Prokaryotic Methylation
dcm
Cytosine methylase mutation—methylates the C5 position of the
internal cytosine residue in the sequence 5′...CCTGG...3′ or 5′...
CCAGG...3′.
dam
Adenine methylase mutation—methylates the N6 position of the
adenine residue in the sequence 5′....GATC...3′.
Eukaryotic Methylation
CpG
Methylates the C5 position of the cytosine residue in the
dinucleotide recognition sequence 5′...CG...3′.
CpNpG Methylates the C5 position of the cytosine residue in the
trinucleotide recognition sequence 5′...CNG...3′ (N = any base).
Enzyme AatII
AccB7I
AccIII
Acc65I
ApaI
AvaI
AvaII
BalI
BamHI
BanII
BbuI
BclI
BglI
BglII
Bsp1286I
BssHII
BstEII
BstOI
BstXI
BstZI
CfoI
ClaI
CspI
Csp45I
DdeI
Eco47III
EcoRI
HaeIII
HhaI
HincII
HindIII
HpaII
KpnI
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This table lists the sensitivities of several Promega restriction enzymes to site-specific methylation at dam, dcm, CpG and CpNpG sites (p = phos­phoryl group).
These four modifications are frequently found in DNA of bacteria, eukaryotes or their viruses. Many strains of E. coli contain the site-specific dam and dcm
DNA methylases. Higher eukaryotes contain the site-specific CpG and CpNpG DNA methylases. In mammalian genomes, methylation occurs mainly at the CG
dinucleotide. In plant genomes, methylation may occur at both the CG and CNG sequences.
Recognition Sequence
GACGTC
CCANNNNNTGG
TCCGGA
GGTACC
GGGCCC
CYCGRG
GGWCC
TGGCCA
GGATCC
GRGCYC
GCATGC
TGATCA
GCCNNNNNGGC
AGATCT
GDGCHC
GCGCGC
GGTNACC
CCWGG
CCANNNNNNTGG
CGGCCG
GCGC
ATCGAT
CGGWCCG
TTCGAA
CTNAG
AGCGCT
GAATTC
GGCC
GCGC
GTYRAC
AAGCTT
CCGG
GGTACC
dam
i
i
s(ol)
i
i
i
i
i
i
i
i
s
i
i
i
i
i
i
i
i
i
s(ol)
i
i
i
i
i
i
i
i
i
i
i
dcm
i
s(ol)
i
s(ol)
s(ol)
i
s(ol)
s(ol)
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
CpG CpNpG
s
i
i
i
i
i
i
i
s(ol)
i
s
i
s(ol) s(ol)
i
s(ol)
i
s(ol)
i
i
i
i
i
i
s(ol) s(ol)
i
s(ol)
i
i
s
i
i
i
i
n.d.
i
i
s(ol) s(ol)
s
n.d.
s
i
i
s
s
i
i
s(ol)
s
i
s(ol)
i
i
s(ol)
s
s(ol)
i
i
i
i
s
s
i
i
For further information regarding site-specific methylation, refer to
McClelland, M., Nelson, M. and Raschke, E. (1994) Nucl. Acids Res. 22,
3640.
Key:
s = i=
s(ol) =
n.d. =
sensitive to this methylation
insensitive to this methylation
overlapping (sensitive when restriction site overlaps methylation sequence)
information not available
Enzyme MboII
MluI
MspI
NaeI
NarI
NdeII
NheI
NotI
NruI PmeI
PstI
PvuI
PvuII
SacI
SacII
SalI
Sau3AI
ScaI
SfiI
SgfI
SinI
SmaI
SnaBI
SphI
StuI
TaqI
XbaI
XhoI
XhoII
XmaI
XmnI
Recognition Sequence
GAAGA(8/7)
ACGCGT
CCGG
GCCGGC
GGCGCC
GATC
GCTAGC
GCGGCCGC
TCGCGA
GTTTAAAC
CTGCAG
CGATCG
CAGCTG
GAGCTC
CCGCGG
GTCGAC
GATC
AGTACT
GGCCNNNNNGGCC
GCGATCGC
GGWCC
CCCGGG
TACGTA
GCATGC
AGGCCT
TCGA
TCTAGA
CTCGAG
RGATCY
CCCGGG
GAANNNN
dam
s(ol)
i
i
i
i
s
i
i
s(ol)
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
s(ol)
s(ol)
i
i
i
i
dcm
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
i
s(ol)
i
i
i
i
i
s(ol)
i
i
i
i
i
i
* PmeI is inhibited by methylation of cytosine residues on both strands.
CpG CpNpG
i
i
s
i
i
s
s
s
s
i
i
i
s(ol) s(ol)
s
s
s
i
s(ol)* s(ol)*
i
s
s
s(ol)
i
s
i
i
s
s
s
n.d.
s(ol) s(ol)
i
i
s(ol) s(ol)
s
n.d.
i
s(ol)
s
s
s
i
i
i
i
s(ol)
i
i
i
i
s
i
i
s(ol)
i
n.d.
n.d. n.d.