Restriction Enzymes

Enzyme AccI AatII

Recognition 5' GTMKAC 5' GACGTC
Sequence: 3' CAKMTG 3' CTGCAG

Restriction site: 5' ---GT   MKAC--- 3' 5' ---GACGT   C--- 3'

3' ---CAKM   TG--- 5' 3' ---C   TGCAG--- 5'
Isoschizomers FblI, XmiI Ssp5230I, ZraI

Source Acinettobacter calcoaceticus Acetobacter aceti

Dam Sensitivity Not sensitive Not sensitive

Dcm Sensitivity Not sensitive Not sensitive

Methylase influencin CG methylase (5mCG) CG methylase (5mCG)

g restriction cleavage
efficiency and its
recognition sequence

Cleavage sites GTMKA5mCG GA5mCGTC

blocked by GT5mCGAC
methylation

Sequences blocked  GTMK6mAC GA5mCGTC

by methylation GTMKA5mC3 GACGT5mC
GT5mCGAC4
Buffer 50% activity in NEBuffer 1  100% activity in CutSmart® Buffer
Star status activity Cut in the presence of  unrelated site may often be cut in
NEBuffers presence of DMSO
Enzyme activity units Number of units required to  4–12 units/µl
cleave 1 µg of supercoiled
plasmid DNA in one hour: 4
units
Genome DNA Staphylococcus aureus genome Staphylococcus aureus genomic
analysis DNA DNA

References:

 https://www.takarabio.com/learning-centers/cloning/traditional-molecular-
cloning/restriction-enzyme-overview/restriction-enzymes-affected-by-methylation
 https://www.takarabio.com/products/cloning/restriction-enzymes/aatii
 https://www.takarabio.com/products/cloning/restriction-enzymes/acci
 https://international.neb.com/tools-and-resources/selection-charts/dam-dcm-and-cpg-
methylation