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duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
linear plasmid DNA pDRM-2R + ATP
?
-
-
-
?
plasmid DNA + H2O
?
-
EcoKI prefers to have a partially filled DNA-binding site rather than one fully occupied by non-specific DNA. Dimerization of EcoKI does not occur before DNA binding and takes place on specific sites before any looping. Dimerization occurs before the two specific sites are bought together. Looping initially occurs between a target site and a non-specific region of DNA
-
-
?
plasmid pTK-neo + ATP
?
-
the enzyme recognises the symmetrical sequence GAAN7TTC at position 2535 bp
-
-
?
plasmid pUC19 + ATP
?
-
the enzyme recognises the symmetrical sequence GAAN7TTC at positions 1126 bp and 2294 bp
-
-
?
supercoiled plasmid DNA pRK + ATP
?
-
-
-
?
synthetic oligonucleotide + ATP
?
additional information
?
-
DNA + H2O
?
-
EcoR124I couples ATP hydrolysis to bidirectional DNA translocation
-
-
?
DNA + H2O
?
-
the enzyme must overcome a similar slow step before translocation reaches a steady state
-
-
?
DNA + H2O
?
-
HsdR subunit can produce only a single cleavage of the phosphodiester backbone of the DNA but can cooperate with another HsdR subunit to produce full DNA cleavage, producing DNA with overhanging ends of single-stranded DNA. On a single-site plasmid, cleavage requires the association of HsdR, from solution, to the cleavage complex in order to produce double-strand cleavage
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
-
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
overview of recognition sequences
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
the wild type enzyme EcoK cleaves circular DNA. Only one endonuclease molecule is required per cleavage event. Cleavage of linear DNA may require a second endonuclease molecule
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
the enzyme is both a restriction endonuclease and a modification methylase. Hemi-methylated DNA is the preferred substrate for methylation
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
the site of restriction cleavage is random, occuring between 1 and 5 kb from the recognition site. The modification methylase acts directly at the recognition sequence
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
the modification methylase binds sequence specifically to DNA and protects a 25bp fragment containing its cognate recognition sequence from digestion by exonuclease III, specific adenine on each strand of DNA is the site of methylation
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
supercoiled with one or two SR124I recognition sites is cleaved by the same mechanism. Nicked-circle DNA is an intermediate of the cleavage reaction
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
EcoAI preferentially generates 3'-overhangs of 2-3 nt. Displays some preference for the formation of 5'-overhangs of a length of 6-7 and 3-5 nt, respectively. type I restriction enzymes require two restriction subunits to introduce DNA double-stran breaks, each providing one catalytic center for phosphodiester bond hydrolysis
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
EcoKI displays some preference for the formation of 5'-overhangs of a length of 6-7 and 3-5 nt, respectively. Type I restriction enzymes require two restriction subunits to introduce DNA double-stran breaks, each providing one catalytic center for phosphodiester bond hydrolysis
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
EcoR124I displays some preference for the formation of 5'-overhangs of a length of 6-7 and 3-5 nt, respectively. Type I restriction enzymes require two restriction subunits to introduce DNA double-stran breaks, each providing one catalytic center for phosphodiester bond hydrolysis
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
initiation of translocation by type I restriction-modification enzymes is associated with a short DNA extrusion
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
the two motor subunits of Eco124I are independent motors that translocate along the helical pitch of the DNA. Dynamic termination and reinitiation of translocation activity is governed by disassembly and reassembly of the enzyme
-
-
?
duplex DNA + ATP
double-stranded DNA fragments with terminal 5'-phosphate + ADP + inorganic phosphate
-
type I enzymes recognize bipartite DNA sequences comprising two half-sequences separated by a gap, for example, AACNNNNNNGTGC (AAC N6 GTGC) where N=any base
-
-
?
synthetic oligonucleotide + ATP
?
-
recognition sequence: CA(underlined)C(5N)T(underlined)GGC
-
-
?
synthetic oligonucleotide + ATP
?
-
recognition sequence: GA(underlined)C(5N)RT(underlined)AAY
-
-
?
synthetic oligonucleotide + ATP
?
-
recognition sequence: GCA(underlined)(6N)CT(underlined)GA
-
-
?
synthetic oligonucleotide + ATP
?
-
recognition sequence: GTCA(underlined)(6N)T(underlined)GAY
-
-
?
additional information
?
-
-
-
-
-
?
additional information
?
-
-
the database REBASE contains information about recognition sites and cleavage sites
-
-
?
additional information
?
-
-
a bacterial population may switch the recognition sequence of its type I restriction-modification system by single recombination events and thus is able to maintain a prokaryotic analogue of the immune system of variable specificity
-
-
?
additional information
?
-
-
enzyme restricts the exchange of genetic material between bacteria of different strains or species
-
-
?
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116000
-
x * 116000, HsdR, x * 55000, HsdM, x * 43000, HsdS
130000
1 * 94000, dynamic light scattering, 1 * 130000, analytical ultracentrifugation, subunit HsdR, subunit is globular and fairly compact
290000 - 315000
-
pentameric enzyme form R2M2S1, gel filtration
400000
-
EcoK, sucrose density gradient sedimentation
43000
-
x * 116000, HsdR, x * 55000, HsdM, x * 43000, HsdS
431000
-
x * 431000, calculated from amino acid sequence
52000
-
alpha2,beta2,gamma1, 2 * 135000 + 2 * 62000 + 1 * 52000, EcoK, SDS-PAGE
60000
-
alpha2,beta4,gamma2, 2 * 135000 + 4 * 60000 + 2 * 55000, EcoB, SDS-PAGE
62000
-
alpha2,beta2,gamma1, 2 * 135000 + 2 * 62000 + 1 * 52000, EcoK, SDS-PAGE
94000
1 * 94000, dynamic light scattering, 1 * 130000, analytical ultracentrifugation, subunit HsdR, subunit is globular and fairly compact
135000
-
alpha2,beta2,gamma1, 2 * 135000 + 2 * 62000 + 1 * 52000, EcoK, SDS-PAGE
135000
-
alpha2,beta4,gamma2, 2 * 135000 + 4 * 60000 + 2 * 55000, EcoB, SDS-PAGE
55000
-
alpha2,beta4,gamma2, 2 * 135000 + 4 * 60000 + 2 * 55000, EcoB, SDS-PAGE
55000
-
x * 116000, HsdR, x * 55000, HsdM, x * 43000, HsdS
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