2.7.7.B22: transposase
This is an abbreviated version!
For detailed information about transposase, go to the full flat file.
Word Map on EC 2.7.7.B22
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2.7.7.B22
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transposition
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transposable
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drosophila
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chromatin
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strand
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melanogaster
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cassette
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phage
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hyperactive
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germline
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transgenesis
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imperfect
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protein-dna
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atac-seq
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helper
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cointegrate
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nonautonomous
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brucellae
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brucellosis
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resolvase
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retroviral
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bacteriophage
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footprint
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musca
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burnetii
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selfish
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medicine
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remobilization
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reinsertion
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extrachromosomal
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coxiella
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p-elements
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medaka
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biotechnology
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thrombectomy
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trichoplusia
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mobilizable
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recombinases
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nonviral
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biovars
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molecular biology
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melitensis
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helix-turn-helix
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analysis
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irl
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excise
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dysgenesis
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abortus
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integrons
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prophage
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mite
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retrotransposons
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subterminal
- 2.7.7.B22
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transposition
-
transposable
- drosophila
- chromatin
- strand
- melanogaster
- cassette
- phage
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hyperactive
-
germline
-
transgenesis
-
imperfect
-
protein-dna
-
atac-seq
-
helper
-
cointegrate
-
nonautonomous
-
brucellae
- brucellosis
- resolvase
-
retroviral
- bacteriophage
-
footprint
-
musca
- burnetii
-
selfish
- medicine
-
remobilization
-
reinsertion
-
extrachromosomal
- coxiella
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p-elements
- medaka
- biotechnology
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thrombectomy
-
trichoplusia
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mobilizable
-
recombinases
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nonviral
-
biovars
- molecular biology
- melitensis
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helix-turn-helix
- analysis
- irl
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excise
- dysgenesis
- abortus
- integrons
- prophage
- mite
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retrotransposons
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subterminal
Reaction
binds to the ends of a transposon (DNA that moves) and catalyzes the movement of the transposon to another part of the genome by a cut and paste mechanism or a replicative transposition mechanism =
Synonyms
Ac transposase, AcTPase, APM_2825, Buster, DbuzGalileo, DDE-type integrase/transposase/recombinase, DDE_Tnp_1_7 domain-containing protein, DNA transposase, ECO103_3558, Galileo transposase, hAT transposase, hAT-transposase 1, Hermes, Hermes transposase, Hsmar1 transposase, ICE6013, IS10 transposase, IS1341-type transposase, IS1634 transposase, IS200 transposase, IS30-like transposase, IS608, IS711, ISC1041, Kat1, maize activator transposase, mariner transposase, Mbar_A1742, PGM, piggyBac transposase, PiggyMac, Rev 1 transposase, Rev 2 transposase, SB transposase, SB100X, SETMAR, sleeping beauty transposase, SSO1474, Ta0474, Tgf2 TPase, Tgf2 transposase, Tgf2TPase, Tn4430 TnpA, Tn5, Tn5 transposase, Tn7 transposase, TnpA, TnpA 155-amino acid transposase, TnpA transposase, TnsA, TnsB, Tol2, Tpase, transposase for transposon Tn4430, transposase for transposon Tn5, wbk IS transposase, wbkA flanking transposase, wbkA-flanking IS
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2.7.7.B22 transposaseAdvanced search results
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results (Engineering
Engineering on EC 2.7.7.B22 - transposase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
A174V
substitution close to the putative N-terminal DNA-binding domain of TnpA, reduces immunity by approximately sixfold. Transposition activity is comparable to that of wild-type
E740G
C-terminal substitutions within the predicted RNaseH fold, reduces immunity up to about12fold. Transposition activity is comparable to that of wild-type
S911R
C-terminal substitution adjacent to the predicted RNaseH fold, reduces immunity up to about 25fold. Transposition activity is comparable to that of wild-type
W24R
substitution within the putative N-terminal DNA-binding domain of TnpA, reduces immunity by approximately twofold. Transposition activity is comparable to that of wild-type
W24R/A174V/E740G
triple mutant is hyperactive in vivo, giving elevated levels of transposition into both permissive and immune targets
K212R/P214R/G251R/A338V
i.e. Tn5-059, mutant displays a lowered GC insertion bias. Tn5-059 reduces AT dropout and increases uniformity of genome coverage in both bacterial genomes and human genome
K339
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site-directed mutagenesis, the mutant shows 20% reduced activity compared to the wild-type enzyme
N280
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site-directed mutagenesis, the mutant shows 50% reduced activity compared to the wild-type enzyme
C130A/C133A
C402A/H405A
W576A
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mutation predicted to be impaired for hairpin formation, mutant is active for DNA cleavage and supports wild type levels of mating-type switching
K569A
mutant exhibits a sharp decrease in its apparent binding affinity to LE1-35 substrate
R567A
mutant exhibits a sharp decrease in its apparent binding affinity to LE1-35 substrate
Y558A
mutant exhibits a decrease in its apparent binding affinity to LE1-35 substrate
additional information
C130A/C133A
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mutations in the BED zinc-finger domain, result in an unspecific nuclease activity
C402A/H405A
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mutant binds DNA normally and is critical for hairpin formation
C402A/H405A
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mutant completely blocks hairpinning and switching, but still generates nicks in the DNA
additional information
compared with wild-type TnpA, Tn4430 TnpA mutants that are proficient in transposition but impaired in target immunity exhibit deregulated activities. They spontaneously assemble a unique asymmetric synaptic complex in which one TnpA molecule simultaneously binds two transposon ends. In this complex, TnpA is in an activated state competent for DNA cleavage and strand transfer. Wild-type TnpA can form this complex only on precleaved ends mimicking the initial step of transposition
additional information
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generation of strain Rev2, by Rev 1 double in-frame deletion mutation in ISBm-1 transposase and GI-2 phage integrase giving mutant strain Rev2, improves the stability of Brucella melitensis Rev 1 vaccine. The parental Rev 1 strain, the Rev 2 double mutant DELTAISBm1DELTAint strain, and the virulent Bacillus melitensis strain H38 (as a control) are inoculated
additional information
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generation of strain Rev2, by Rev 1 double in-frame deletion mutation in ISBm-1 transposase and GI-2 phage integrase giving mutant strain Rev2, improves the stability of Brucella melitensis Rev 1 vaccine. The parental Rev 1 strain, the Rev 2 double mutant DELTAISBm1DELTAint strain, and the virulent Bacillus melitensis strain H38 (as a control) are inoculated
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additional information
construction of C-terminal deletion mutants, i.e. pEGFP-C1-Tgf2TPDELTA31C containing a 31-aa C-terminal deletion of Tgf2 transposase that included the predicted 15-amino acid NLS, pEGFP-C1-Tgf2TPDELTA120N containing a 120-aa N-terminal deletion of Tgf2 transposase, and pEGFP-C1-Tgf2TPDELTA16C containing a 16-aa C-terminal deletion of Tgf2 transposase. Loss of the nuclear localization signal (NLS) domain results in expression in the cytoplasm but not in the nucleus
additional information
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construction of C-terminal deletion mutants, i.e. pEGFP-C1-Tgf2TPDELTA31C containing a 31-aa C-terminal deletion of Tgf2 transposase that included the predicted 15-amino acid NLS, pEGFP-C1-Tgf2TPDELTA120N containing a 120-aa N-terminal deletion of Tgf2 transposase, and pEGFP-C1-Tgf2TPDELTA16C containing a 16-aa C-terminal deletion of Tgf2 transposase. Loss of the nuclear localization signal (NLS) domain results in expression in the cytoplasm but not in the nucleus
additional information
contruction of two truncated recombinant Tgf2 transposases with deletions in the N-terminal zinc finger domain, S1- and S2-Tgf2TPase, from goldfish cDNAs. Both truncated Tgf2TPases lost their DNA-binding ability in vitro, specifically at the ends of Tgf2 transposon than native L-Tgf2TPase. Mutant S1- and S2-Tgf2TPases mediate gene transfer in the zebrafish genome in vivo at a significantly lower efficiency (21%-25%), in comparison with L-Tgf2TPase (56% efficiency). Compared to L-Tgf2TPase, truncated Tgf2TPases catalyze imprecise excisions with partial deletion of TE ends and/or plasmid backbone insertion/deletion. The gene integration into the zebrafish genome mediated by truncated Tgf2TPases is imperfect, creating incomplete 8-bp target site duplications at the insertion sites
additional information
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contruction of two truncated recombinant Tgf2 transposases with deletions in the N-terminal zinc finger domain, S1- and S2-Tgf2TPase, from goldfish cDNAs. Both truncated Tgf2TPases lost their DNA-binding ability in vitro, specifically at the ends of Tgf2 transposon than native L-Tgf2TPase. Mutant S1- and S2-Tgf2TPases mediate gene transfer in the zebrafish genome in vivo at a significantly lower efficiency (21%-25%), in comparison with L-Tgf2TPase (56% efficiency). Compared to L-Tgf2TPase, truncated Tgf2TPases catalyze imprecise excisions with partial deletion of TE ends and/or plasmid backbone insertion/deletion. The gene integration into the zebrafish genome mediated by truncated Tgf2TPases is imperfect, creating incomplete 8-bp target site duplications at the insertion sites
additional information
P13988; P13989
construction of diverse truncation mutants of TnsB, TnsC, and TnsD, and of double-mutant MBP-TnsC361555 P468A/L470A
additional information
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development of a simple and precise method for genome manipulation in Escherichia coli that alters the gene sequence without leaving foreign DNA in the chromosome. This strategy involves PCR amplification of a DNA cassette containing ISHp608-LE (left end)-antibiotic resistance gene-counterselection marker-ISHp608-RE (right end) by using primers containing extensions homologous to the adjacent regions of the target gene on the chromosome. The lambda Red-mediated recombination of the PCR product and antibiotic resistance screening results in transformants with a modified gene target. The ISHp608-LE-antibiotic resistance gene-counterselection marker-ISHp608-RE cassette can then be excised using a temperature sensitive plasmid expressing the TnpA transposase, which precisely cleaves ISHp608-LE and ISHp608-RE without leaving a scar sequence. For introduction of IS608 LE and RE into the gene of interest, lambda-Red recombination is utilized, which does not require in vitro manipulations such as restriction digestion, ligation or construction of a suicide vector. Diagram of plasmids containing selectable and excisable IS608 cassettes, overview
additional information
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development of a simple and precise method for genome manipulation in Escherichia coli that alters the gene sequence without leaving foreign DNA in the chromosome. This strategy involves PCR amplification of a DNA cassette containing ISHp608-LE (left end)-antibiotic resistance gene-counterselection marker-ISHp608-RE (right end) by using primers containing extensions homologous to the adjacent regions of the target gene on the chromosome. The lambda Red-mediated recombination of the PCR product and antibiotic resistance screening results in transformants with a modified gene target. The ISHp608-LE-antibiotic resistance gene-counterselection marker-ISHp608-RE cassette can then be excised using a temperature sensitive plasmid expressing the TnpA transposase, which precisely cleaves ISHp608-LE and ISHp608-RE without leaving a scar sequence. For introduction of IS608 LE and RE into the gene of interest, lambda-Red recombination is utilized, which does not require in vitro manipulations such as restriction digestion, ligation or construction of a suicide vector. Diagram of plasmids containing selectable and excisable IS608 cassettes, overview
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additional information
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design of two additional hyperactive transposase variants using the enzyme crystal structure, evaluation of structure-based engineering of tailored SB transposases, overview
additional information
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introduction of mutation I212S to variant SB100X increases solubility. Additional substitution C176S generates a remarkably high solubility. Mutations C197S, C304S and C316S compromise protein solubility
additional information
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mutations of predicted catalytic residues abolish both DNA cleavage and strand-transfer. Kat1 mutants defective for cleavage in vitro are also defective for mating-type switching
additional information
introduction of three point mutations into the interface disrupting the octamer shows that the resulting dimers are catalytically active in vitro. Generation of dimers by deleting the helix and surrounding residues, HermesDELTA497-516, also leads to active dimers in vitro, that can catalyze all of the catalytic steps. Although Hermes dimers are hyperactive in vitro at low ionic strength, their activities are severely reduced under more physiologically relevant conditions. Hermes dimers are inactive in vivo
additional information
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introduction of three point mutations into the interface disrupting the octamer shows that the resulting dimers are catalytically active in vitro. Generation of dimers by deleting the helix and surrounding residues, HermesDELTA497-516, also leads to active dimers in vitro, that can catalyze all of the catalytic steps. Although Hermes dimers are hyperactive in vitro at low ionic strength, their activities are severely reduced under more physiologically relevant conditions. Hermes dimers are inactive in vivo
