BRENDA - Enzyme Database show
show all sequences of 2.7.1.145

Identification of residues involved in the specificity and regulation of the highly efficient multisubstrate deoxyribonucleoside kinase from Drosophila melanogaster

Knecht, W.; Munch-Petersen, B.; Piskur, J.; J. Mol. Biol. 301, 827-837 (2000)

Data extracted from this reference:

Engineering
Amino acid exchange
Commentary
Organism
I102M/N117S/M118V/D208N
clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine
Drosophila melanogaster
I47T/N210D
clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine
Drosophila melanogaster
M1T/T85A/N121S
clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine and 2',3'-dideoxycytidine
Drosophila melanogaster
N210D/L239P
clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine
Drosophila melanogaster
N38D/N64D
clones harbouring the mutant enzyme are more sensitive specifically to 3'-azido-2',3'-dideoxythymidine
Drosophila melanogaster
N45D/N64D
clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and other nucleoside analogs; mutant enzyme with the largest und universal increase in sensitivity of all mutants tested. 316fold increase to 3'-azido-2',3'-dideoxythymidine, more than 11fold to 2',3'-dideoxycytidine, and 3.2fold increase in sensitivity to beta-D-arabinofuranosylcytosine and 2',3'-dideoxyadenosine. Higher Km-values for native substrates than wild-type enzyme and Vmax-values are substantially lower, decrease in feedback inhibition by TTP, Km and Vmax values for 3'-azido-2',3'-dideoxythymidine and Km value for 2',3'-dideoxycytidine are nearly unchanged
Drosophila melanogaster
N59D/M118V/Y179H
clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine
Drosophila melanogaster
N64S/L68S/M69L
clones harbouring the mutant enzyme are more sensitive specifically to 3'-azido-2',3'-dideoxythymidine
Drosophila melanogaster
T12A/V84A/N213S
clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine
Drosophila melanogaster
T85A
clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine and 2',3'-dideoxycytidine
Drosophila melanogaster
V84A
clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine
Drosophila melanogaster
Inhibitors
Inhibitors
Commentary
Organism
Structure
TTP
mutant enzyme N45D/N64D shows a decrease in feedback inhibition compared to the wild-type enzyme
Drosophila melanogaster
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0072
-
3'-azido-2',3'-dideoxythymidine
mutant enzyme N45D/N64D
Drosophila melanogaster
0.0083
-
3'-azido-2',3'-dideoxythymidine
-
Drosophila melanogaster
0.0242
-
thymidine
mutant enzyme N45D/N64D
Drosophila melanogaster
0.0964
-
2'-deoxycytidine
mutant enzyme N45D/N64D
Drosophila melanogaster
1.093
-
2',3'-dideoxycytidine
mutant enzyme N45D/N64D
Drosophila melanogaster
1.124
-
2',3'-dideoxycytidine
-
Drosophila melanogaster
2.004
-
2'-deoxyguanosine
mutant enzyme N45D/N64D
Drosophila melanogaster
3.16
-
2'-deoxyadenosine
mutant enzyme N45D/N64D
Drosophila melanogaster
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Drosophila melanogaster
-
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(E)-5-(2-bromovinyl)-2'-deoxyuridine + ATP
54% of the activity with thymidine
636844
Drosophila melanogaster
(E)-5-(2-bromovinyl)-2'-deoxyuridine 5'-phosphate + ADP
-
-
-
?
1-beta-D-arabinofuranosylcytosine + ATP
58.2% of the activity with thymidine
636844
Drosophila melanogaster
1-beta-D-arabinofuranosylcytosine 5'-phosphate + ADP
-
-
-
?
1-beta-D-arabinofuranosylthymine + ATP
53.9% of the activity with thymidine
636844
Drosophila melanogaster
1-beta-D-arabinofuranosylthymine 5'-phosphate + ADP
-
-
-
?
2',3'-dideoxyadenosine + ATP
0.9% of the activity with thymidine
636844
Drosophila melanogaster
2',3'-dideoxyadenosine 5'-phosphate + ADP
-
-
-
?
2',3'-dideoxycytidine + ATP
-
636844
Drosophila melanogaster
2',3'-dideoxycytidine 5'-phosphate + ADP
-
-
-
?
2'-deoxyadenosine + ATP
-
636844
Drosophila melanogaster
2'-deoxyadenosine 5'-phosphate + ADP
-
-
-
?
2'-deoxycytidine + ATP
-
636844
Drosophila melanogaster
2'-deoxycytidine 5'-phosphate + ADP
-
-
-
?
2'-deoxyguanosine + ATP
-
636844
Drosophila melanogaster
2'-deoxyguanosine 5'-phosphate + ADP
-
-
-
?
2'-deoxyuridine + ATP
113% of the activity with thymidine
636844
Drosophila melanogaster
2'-deoxyuridine 5'-phosphate + ADP
-
-
-
?
2-chloro-2'-deoxyadenosine + ATP
126% of the activity with thymidine
636844
Drosophila melanogaster
2-chloro-2'-deoxyadenosine 5'-phosphate + ADP
-
-
-
?
3'-azido-2',3'-dideoxythymidine + ATP
-
636844
Drosophila melanogaster
3'-azido-2',3'-dideoxythymidine 5'-phosphate + ADP
-
-
-
?
5-fluoro-2'-deoxyuridine + ATP
92% of the activity with thymidine
636844
Drosophila melanogaster
5-fluoro-2'-deoxyuridine 5'-phosphate + ADP
-
-
-
?
adenosine + ATP
0.82% of the activity with thymidine
636844
Drosophila melanogaster
adenosine 5'-phosphate + ADP
-
-
-
?
cytidine + ATP
16% of the activity with thymidine
636844
Drosophila melanogaster
cytidine 5'-phosphate + ADP
-
-
-
?
guanosine + ATP
1.18% of the activity with thymidine
636844
Drosophila melanogaster
guanosine 5'-phosphate + ADP
-
-
-
?
thymidine + ATP
-
636844
Drosophila melanogaster
thymidine 5'-phosphate + ADP
-
-
-
?
uridine + ATP
6.45% of the activity with thymidine
636844
Drosophila melanogaster
uridine 5'-phosphate + ADP
-
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.036
-
3'-azido-2',3'-dideoxythymidine
-
Drosophila melanogaster
0.052
-
3'-azido-2',3'-dideoxythymidine
mutant enzyme N45D/N64D
Drosophila melanogaster
0.076
-
2'-deoxyguanosine
mutant enzyme N45D/N64D
Drosophila melanogaster
0.54
-
2',3'-dideoxycytidine
mutant enzyme N45D/N64D
Drosophila melanogaster
0.828
-
2'-deoxyadenosine
mutant enzyme N45D/N64D
Drosophila melanogaster
1.22
-
thymidine
mutant enzyme N45D/N64D
Drosophila melanogaster
4.03
-
2'-deoxycytidine
mutant enzyme N45D/N64D
Drosophila melanogaster
4.2
-
2',3'-dideoxycytidine
-
Drosophila melanogaster
14.2
-
thymidine
pH 7.8, 22°C, recombinant enzyme
Drosophila melanogaster
15.1
-
2'-deoxyguanosine
pH 7.8, 22°C, recombinant enzyme
Drosophila melanogaster
16.5
-
2'-deoxycytidine
pH 7.8, 22°C, recombinant enzyme
Drosophila melanogaster
20.6
-
2'-deoxyadenosine
pH 7.8, 22°C, recombinant enzyme
Drosophila melanogaster
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
I102M/N117S/M118V/D208N
clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine
Drosophila melanogaster
I47T/N210D
clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine
Drosophila melanogaster
M1T/T85A/N121S
clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine and 2',3'-dideoxycytidine
Drosophila melanogaster
N210D/L239P
clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine
Drosophila melanogaster
N38D/N64D
clones harbouring the mutant enzyme are more sensitive specifically to 3'-azido-2',3'-dideoxythymidine
Drosophila melanogaster
N45D/N64D
clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and other nucleoside analogs; mutant enzyme with the largest und universal increase in sensitivity of all mutants tested. 316fold increase to 3'-azido-2',3'-dideoxythymidine, more than 11fold to 2',3'-dideoxycytidine, and 3.2fold increase in sensitivity to beta-D-arabinofuranosylcytosine and 2',3'-dideoxyadenosine. Higher Km-values for native substrates than wild-type enzyme and Vmax-values are substantially lower, decrease in feedback inhibition by TTP, Km and Vmax values for 3'-azido-2',3'-dideoxythymidine and Km value for 2',3'-dideoxycytidine are nearly unchanged
Drosophila melanogaster
N59D/M118V/Y179H
clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine
Drosophila melanogaster
N64S/L68S/M69L
clones harbouring the mutant enzyme are more sensitive specifically to 3'-azido-2',3'-dideoxythymidine
Drosophila melanogaster
T12A/V84A/N213S
clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine
Drosophila melanogaster
T85A
clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine and 2',3'-dideoxycytidine
Drosophila melanogaster
V84A
clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine
Drosophila melanogaster
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
TTP
mutant enzyme N45D/N64D shows a decrease in feedback inhibition compared to the wild-type enzyme
Drosophila melanogaster
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0072
-
3'-azido-2',3'-dideoxythymidine
mutant enzyme N45D/N64D
Drosophila melanogaster
0.0083
-
3'-azido-2',3'-dideoxythymidine
-
Drosophila melanogaster
0.0242
-
thymidine
mutant enzyme N45D/N64D
Drosophila melanogaster
0.0964
-
2'-deoxycytidine
mutant enzyme N45D/N64D
Drosophila melanogaster
1.093
-
2',3'-dideoxycytidine
mutant enzyme N45D/N64D
Drosophila melanogaster
1.124
-
2',3'-dideoxycytidine
-
Drosophila melanogaster
2.004
-
2'-deoxyguanosine
mutant enzyme N45D/N64D
Drosophila melanogaster
3.16
-
2'-deoxyadenosine
mutant enzyme N45D/N64D
Drosophila melanogaster
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(E)-5-(2-bromovinyl)-2'-deoxyuridine + ATP
54% of the activity with thymidine
636844
Drosophila melanogaster
(E)-5-(2-bromovinyl)-2'-deoxyuridine 5'-phosphate + ADP
-
-
-
?
1-beta-D-arabinofuranosylcytosine + ATP
58.2% of the activity with thymidine
636844
Drosophila melanogaster
1-beta-D-arabinofuranosylcytosine 5'-phosphate + ADP
-
-
-
?
1-beta-D-arabinofuranosylthymine + ATP
53.9% of the activity with thymidine
636844
Drosophila melanogaster
1-beta-D-arabinofuranosylthymine 5'-phosphate + ADP
-
-
-
?
2',3'-dideoxyadenosine + ATP
0.9% of the activity with thymidine
636844
Drosophila melanogaster
2',3'-dideoxyadenosine 5'-phosphate + ADP
-
-
-
?
2',3'-dideoxycytidine + ATP
-
636844
Drosophila melanogaster
2',3'-dideoxycytidine 5'-phosphate + ADP
-
-
-
?
2'-deoxyadenosine + ATP
-
636844
Drosophila melanogaster
2'-deoxyadenosine 5'-phosphate + ADP
-
-
-
?
2'-deoxycytidine + ATP
-
636844
Drosophila melanogaster
2'-deoxycytidine 5'-phosphate + ADP
-
-
-
?
2'-deoxyguanosine + ATP
-
636844
Drosophila melanogaster
2'-deoxyguanosine 5'-phosphate + ADP
-
-
-
?
2'-deoxyuridine + ATP
113% of the activity with thymidine
636844
Drosophila melanogaster
2'-deoxyuridine 5'-phosphate + ADP
-
-
-
?
2-chloro-2'-deoxyadenosine + ATP
126% of the activity with thymidine
636844
Drosophila melanogaster
2-chloro-2'-deoxyadenosine 5'-phosphate + ADP
-
-
-
?
3'-azido-2',3'-dideoxythymidine + ATP
-
636844
Drosophila melanogaster
3'-azido-2',3'-dideoxythymidine 5'-phosphate + ADP
-
-
-
?
5-fluoro-2'-deoxyuridine + ATP
92% of the activity with thymidine
636844
Drosophila melanogaster
5-fluoro-2'-deoxyuridine 5'-phosphate + ADP
-
-
-
?
adenosine + ATP
0.82% of the activity with thymidine
636844
Drosophila melanogaster
adenosine 5'-phosphate + ADP
-
-
-
?
cytidine + ATP
16% of the activity with thymidine
636844
Drosophila melanogaster
cytidine 5'-phosphate + ADP
-
-
-
?
guanosine + ATP
1.18% of the activity with thymidine
636844
Drosophila melanogaster
guanosine 5'-phosphate + ADP
-
-
-
?
thymidine + ATP
-
636844
Drosophila melanogaster
thymidine 5'-phosphate + ADP
-
-
-
?
uridine + ATP
6.45% of the activity with thymidine
636844
Drosophila melanogaster
uridine 5'-phosphate + ADP
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.036
-
3'-azido-2',3'-dideoxythymidine
-
Drosophila melanogaster
0.052
-
3'-azido-2',3'-dideoxythymidine
mutant enzyme N45D/N64D
Drosophila melanogaster
0.076
-
2'-deoxyguanosine
mutant enzyme N45D/N64D
Drosophila melanogaster
0.54
-
2',3'-dideoxycytidine
mutant enzyme N45D/N64D
Drosophila melanogaster
0.828
-
2'-deoxyadenosine
mutant enzyme N45D/N64D
Drosophila melanogaster
1.22
-
thymidine
mutant enzyme N45D/N64D
Drosophila melanogaster
4.03
-
2'-deoxycytidine
mutant enzyme N45D/N64D
Drosophila melanogaster
4.2
-
2',3'-dideoxycytidine
-
Drosophila melanogaster
14.2
-
thymidine
pH 7.8, 22°C, recombinant enzyme
Drosophila melanogaster
15.1
-
2'-deoxyguanosine
pH 7.8, 22°C, recombinant enzyme
Drosophila melanogaster
16.5
-
2'-deoxycytidine
pH 7.8, 22°C, recombinant enzyme
Drosophila melanogaster
20.6
-
2'-deoxyadenosine
pH 7.8, 22°C, recombinant enzyme
Drosophila melanogaster
Other publictions for EC 2.7.1.145
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
738739
Kamiya
In vivo selection of active de ...
Drosophila melanogaster
J. Biotechnol.
228
52-57
2016
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1
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1
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1
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1
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3
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1
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3
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1
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1
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1
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1
1
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1
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3
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1
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1
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1
1
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738781
Slot Christiansen
Non-viral deoxyribonucleoside ...
Anopheles gambiae, Bombyx mori, Drosophila melanogaster
J. Genet. Genomics
42
235-248
2015
-
2
-
1
-
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-
1
3
-
18
-
3
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33
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3
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2
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3
1
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18
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33
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7
7
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739211
Jiang
Tanshinone IIA enhances bystan ...
Drosophila melanogaster
Oncol. Rep.
34
1487-1493
2015
-
1
1
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-
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4
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1
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3
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1
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1
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1
1
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4
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1
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1
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3
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1
1
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737404
Serra
-
Immobilized Drosophila melanog ...
Drosophila melanogaster
Adv. Synth. Catal.
356
563-570
2014
-
1
-
-
1
-
-
-
-
1
-
1
-
1
-
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14
-
1
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1
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1
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1
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1
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1
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1
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1
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14
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1
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-
1
-
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-
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-
703396
Betham
Use of Drosophila deoxynucleos ...
Drosophila melanogaster
DNA Repair
9
153-160
2010
-
-
1
-
-
-
-
-
-
-
-
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4
-
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1
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1
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1
1
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1
-
-
-
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1
1
-
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721775
Ito
Cytotoxic effect of Drosophila ...
Drosophila melanogaster
Biol. Pharm. Bull.
33
1223-1227
2010
-
-
1
-
-
-
-
-
-
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5
-
-
-
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1
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702047
Knecht
Drosophila melanogaster deoxyr ...
Drosophila melanogaster
Biochem. Biophys. Res. Commun.
382
430-433
2009
-
-
-
1
-
-
-
1
-
-
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3
-
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1
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1
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1
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1
1
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1
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1
-
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1
-
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-
-
-
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1
1
692289
Mikkelsen
Structural studies of nucleosi ...
Drosophila melanogaster
FEBS J.
275
2151-2160
2008
-
-
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1
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-
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2
-
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1
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Knecht
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Drosophila melanogaster
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Jordheim
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Legent
In vivo analysis of Drosophila ...
Drosophila melanogaster
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676247
Kamiya
Transient expression of Drosop ...
Drosophila melanogaster
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Piskur
Animal deoxyribonucleoside kin ...
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Solaroli
Investigation of the substrate ...
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Krawiec
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Knecht
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Krawiec
Ability of adenosine-2'(3')-de ...
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Eriksson
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A few amino acid substitutions ...
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Johansson
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Munch-Petersen
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Balzarini
The multifunctional deoxynucle ...
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Knecht
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Johansson
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Munch-Petersen
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Munch-Petersen
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