3.1.21.4 D134N/K190N/E204V inactive 730491 3.1.21.4 D140A no detectable activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 D140A/E167A/K169A no detectable activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 D144A no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 D146A no detectable activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 D146A/E177A slight detectable activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 D146A/E177A/K179A no detectable activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 D148A only traces of DNA cleavage activity when used in large excess 666483 3.1.21.4 D148G binds DNA with 2times reduced affinity compared to wild-type protein 666483 3.1.21.4 D148G only traces of DNA cleavage activity when used in large excess 666483 3.1.21.4 D149A no detectable activity in ssDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 D226A no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 D299A no catalytic activity, mutant like wild-type remains in dimeric form 683465 3.1.21.4 D317A 84% of DNA binding compared to wild-type 684042 3.1.21.4 D321A 119% of DNA binding compared to wild-type 684042 3.1.21.4 D328A 76% of DNA binding compared to wild-type 684042 3.1.21.4 D329A 123% of DNA binding compared to wild-type 684042 3.1.21.4 D329A less than 0.01% residual activity, mutant like wild-type remains in dimeric form 683465 3.1.21.4 DELTA362-465 nicking variant of BsaI, nicking occurs on top strand 652938 3.1.21.4 DELTA440-544 nicking variant of BsaI, nicking occurs on bottom strand 652938 3.1.21.4 DELTA446-544 nicking variant of BsaI 652938 3.1.21.4 E139A no detectable activity in ssDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 E147A no detectable activity in ssDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 E155A no detectable activity in ssDNA nicking, low activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 E163A no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 E165A no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 E167A slight activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 E167A/K169A no detectable activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 E167G slight activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 E177A slight activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 E177G no detectable activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 E201A no detectable activity in ssDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 E204Q the mutant possesses 0.004% of wild type activity 730491 3.1.21.4 E211A no detectable activity in dsDNA nicking and no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 E218A no detectable activity in ssDNA nicking, no activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 E227A no detectable activity in dsDNA nicking and no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 E271A no catalytic activity, mutant like wild-type remains in dimeric form 683465 3.1.21.4 E337A less than 0.01% residual activity, mutant like wild-type remains in dimeric form 683465 3.1.21.4 E64A monomer crystallizes in space group C2221, two molecules per asymmetric unit 684038 3.1.21.4 E71A/K190T/E204I inactive 730491 3.1.21.4 E71A/K190T/E204V inactive 730491 3.1.21.4 E71A/S188A/K190T/E204Q inactive 730491 3.1.21.4 E71N/D254G inactive 730491 3.1.21.4 E71Q the mutant possesses 10% of wild type activity 730491 3.1.21.4 E95A no detectable activity in ssDNA and in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 H149L binds DNA with 10times reduced affinity compared to wild-type protein 666483 3.1.21.4 H149L no cleavage activity 666483 3.1.21.4 H291A 96% of DNA binding compared to wild-type 684042 3.1.21.4 H368A 129% of DNA binding compared to wild-type 684042 3.1.21.4 K150R/R236G nicking variant of BsaI, nicking occurs on top strand 652938 3.1.21.4 K169A low activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 K179A no detectable activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit 751757 3.1.21.4 K263A inactive mutant with truncated EcoRII N-domain 693058 3.1.21.4 K324A no catalytic activity, mutant like wild-type remains in dimeric form 683465 3.1.21.4 K328A less than 0.01% residual activity, mutant like wild-type remains in dimeric form 683465 3.1.21.4 L80P the mutant enzyme shows decreased DNA methyltransferase activity at a higher temperature in vivo and in vitro than the wild type enzyme, the activity of the L80P mutant is completely lost at a high temperature -, 692848 3.1.21.4 M357P mutation converts DNA methyltransferase to a type II endonuclease with 5'-CAGCAG-3' restriction site that cleaves only supercoiled DNA but does not act on nicked or linearized DNA 683660 3.1.21.4 additional information a truncated protein comprising the first 259 amino acids of the wild-typedisplays wild-type specificity -, 774908 3.1.21.4 additional information construction of a chimeric endonucelase consisiting of the DNA cleavage domain of isoform BmrI and controller protein C.BclI of the BclI restriction-modification system. The chimeric protein cleaves DNA at specific site in the vicinity of the recognition sequence of C.BclI and requires only half of the C-box sequence for specific cleavage 683940 3.1.21.4 additional information conversion of enzyme from its wild-type homodimeric form into the enzymatically active single-chain variant scPvuII by tandemly joining the two subunits through the peptide linker Gly-Ser-Gly-Gly, crystallization data 683027 3.1.21.4 additional information enzyme is an endonuclease encoded as a protein insert or intein within the yeast V-ATPase catalytic subunit encoding gene vma1. Insertion of the Green Fluorescence Protein into a loop which is located between the endonuclease and splicing domains of the Sce VMA1 intein. The GFP is functional and the additional GFP domain does not prevent intein excision and endonuclease activity. Contrary to wild-type, mutant requires the presence of Mn2+ and not Mg2+ ions for activity 683554 3.1.21.4 additional information evolvement of mutant enzymes with altered DNA cleavage specificities by application of an in vivo positive and negative selection system that applies evolutionary pressure either to favor the cleavage of a desired target sequence or to disfavor the cleavage of a nontarget sequence 683588 3.1.21.4 additional information generation of cleavage specificities of restriction endonucleases by swapping putative target recognition domains between the type IIB enzymes AloI, PpiI from Pseudomonas putida, and TstI from Thermus scotoductus. Individual target recognition domains recognize distinct parts of the bipartite DNA targets of these enzymes and are interchangeable. Engineering of a functional type IIB restriction endonuclease having previously undescribed DNA specificity and application in generation of type II enzymes with predetermined specificity -, 684004 3.1.21.4 additional information mutants of BbvCI with defects in one subunit, either R1-R2+ or R1+R2-, cleave only one strand of the 5'-CC-/-TCAGC-3'/5'-GC-/-TGAGG-3' sequence 666044 3.1.21.4 N138A no detectable activity in ssDNA nicking, mutant R1 subunit/wild-type R2 subunit 751757 3.1.21.4 N308A 104% of DNA binding compared to wild-type 684042 3.1.21.4 N349Y nicking variant of BsmAI, nicking occurs on top strand 652938 3.1.21.4 N415D/R416G variant of BsmAI, both nicking activity and dsDNA cleavage 652938 3.1.21.4 N441D variant of BsaI, cleavage of dsDNA 652938 3.1.21.4 N441D/R442G nicking variant of BsaI, nicking occurs on bottom strand 652938 3.1.21.4 Q115E deprotonated mutant EcoRI is defective in DNA binding at neutral pH 666483 3.1.21.4 Q138F mutation results in a change in the sequence specificity at the center two base pairs of the cognate recognition site. Alteration in preference of HicII for cutting, but not binding, the three cognate sites differening in the center two base pairs.The Q138F HincII/DNA crystal structures show conformational changes in the protein, bound DNA, and at the protein-DNA interface 665724 3.1.21.4 Q175E only traces of DNA cleavage activity when used in large excess 666483 3.1.21.4 R119A subunit BtsIB mutant shows a different digestion pattern from the wild type BtsI. The mutant BtsIB(R119A) acts as a different restriction enzyme with a previously unreported recognition sequence CAGTG(2/0), which is named as BtsI-1 716676 3.1.21.4 R221D nicking variant of BsmAI, nicking on top strand 652938 3.1.21.4 R233D nicking variant of BsmBI, nicking on top strand, about 5-10% cleavage of dsDNA 652938 3.1.21.4 R236D nicking variant of BsaI, nicking on top strand, no cleavage of dsDNA 652938 3.1.21.4 R236G nicking variant of BsaI, 5-10% cleavage of dsDNA 652938 3.1.21.4 R330A inactive mutant with truncated EcoRII C-domain 693058 3.1.21.4 R330A no catalytic activity, mutant like wild-type remains in dimeric form 683465 3.1.21.4 R386S nicking variant of BsaI, nicking occurs on top strand 652938 3.1.21.4 R438D nicking variant of BsmBI, nicking on bottom strand 652938 3.1.21.4 R438G nicking variant of BsmBI 652938 3.1.21.4 R442G nicking variant of BsaI, 5% cleavage of dsDNA 652938 3.1.21.4 R88A crystallization data, autoinhibition/activation mechanism 652931 3.1.21.4 S128L/R236G nicking variant of BsaI, nicking occurs on top strand 652938 3.1.21.4 S188A the mutant possesses 4% of wild type activity 730491 3.1.21.4 S188C the mutant possesses 0.5% of wild type activity 730491 3.1.21.4 W8C/G207E nicking variant of BsaI, nicking occurs on top strand 652938