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3
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0023657032
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(c) Kuchta, R. D.; Mizrahi, V.; Benkovic, P. A.; Johnson, K. A.; Benkovic, S. J. Biochemistry 1987, 26, 8410.
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Kuchta, R.D.1
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Benkovic, S.J.5
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4
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0025736013
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(d)
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(d) Eger, B. T.; Kuchta, R. D.; Carroll, S. S.; Benkovic, P. A.; Dahlberg, M. E.; Joyce, C. M.; Benkovic, S. J. Biochemistry 1991, 30, 1441.
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Eger, B.T.1
Kuchta, R.D.2
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Benkovic, P.A.4
Dahlberg, M.E.5
Joyce, C.M.6
Benkovic, S.J.7
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6
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0026713678
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(f)
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(f) Polesky, A. H.; Dahlberg, M. E.; Benkovic, S. J.; Grindley, N. D. F.; Joyce, C. M. J. Biol. Chem. 1992, 267, 8417.
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Polesky, A.H.1
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Joyce, C.M.5
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10
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84992267203
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note
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-1 (ref 4d).
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13
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0000131255
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(d)
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(d) Fox, J. J.; Von Praag, D.; Wempen, I.; Doerr, I. L.; Cheong, L.; Knoll, J. E.; Eidinoff, M. L.; Bendich, A.; Brown, G. B. J. Am. Chem. Soc. 1959, 81, 178.
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Fox, J.J.1
Von Praag, D.2
Wempen, I.3
Doerr, I.L.4
Cheong, L.5
Knoll, J.E.6
Eidinoff, M.L.7
Bendich, A.8
Brown, G.B.9
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16
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0028291635
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(a)
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(a) Bartholomew, B.; Braun, B. R.; Kassavetis, G. A.; Geiduschek, E. P. J. Biol. Chem. 1994, 269, 18090.
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Bartholomew, B.1
Braun, B.R.2
Kassavetis, G.A.3
Geiduschek, E.P.4
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17
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0029984146
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(b)
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(b) Thorogood, H.; Waters, T. R.; Parker, A. W.; Wharton, C. W.; Connolly, B. A. Biochemistry 1996, 35, 8723.
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Thorogood, H.1
Waters, T.R.2
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Wharton, C.W.4
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18
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0025151421
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(c)
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(c) Newman, P. C.; Nwosu, V. U.; Williams, D. M.; Cosstick, R.; Seela, F.; Connolly, B. A. Biochemistry 1990, 29, 9891.
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Newman, P.C.1
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Connolly, B.A.6
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21
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84992255528
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-
260 nm using the program Oligo (National Biosciences, Plymouth, MN)
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260 nm using the program Oligo (National Biosciences, Plymouth, MN).
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-
-
-
22
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84992245217
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-
The value of Δε was obtained by incubating 25 μM template-primer (DNA-9-mer/DNA-20-mer) and excess 4S-TTP with exonuclease-free Klenow fragment (Unites States Biochemical) under standard buffer conditions (ref. 13) and allowing the reaction to go to completion. Exonuclease-free Klenow fragment was used since with intact Klenow fragment the total absorbance change was smaller and changes in absorbance were observed to reverse direction with prolonged reaction times, indicating slow hydrolysis of the product
-
The value of Δε was obtained by incubating 25 μM template-primer (DNA-9-mer/DNA-20-mer) and excess 4S-TTP with exonuclease-free Klenow fragment (Unites States Biochemical) under standard buffer conditions (ref. 13) and allowing the reaction to go to completion. Exonuclease-free Klenow fragment was used since with intact Klenow fragment the total absorbance change was smaller and changes in absorbance were observed to reverse direction with prolonged reaction times, indicating slow hydrolysis of the product.
-
-
-
-
23
-
-
84992245231
-
-
Note The following three template DNA strands were used: 5′-AAACCCTTGAACGGCTGCGA-OH, 5′-AAACCCTTAAACGGCTGCGA-OH, and 5′-AAACCCTAAAACGGCTGCGA-OH, which when annealed with primer 9-mer in the final template-primer duplex contain sites for incorporation of 2, 3 and 4 (thio)thymidines, respectively, opposite to the underlined A's
-
The following three template DNA strands were used: 5′-AAACCCTTG AA CGGCTGCGA-OH, 5′-AAACCCTT AAA CGGCTGCGA-OH, and 5′-AAACCCT AAAA CGGCTGCGA-OH, which when annealed with primer 9-mer in the final template-primer duplex contain sites for incorporation of 2, 3 and 4 (thio)thymidines, respectively, opposite to the underlined A's.
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-
-
-
25
-
-
84992249669
-
-
2, 1 mM DTT and 0.5 mM EGTA, as used for previous kinetic experiments with Klenow fragment and similar template-primer duplexes (ref 1c). Kinetic parameters (25 °C) were obtained by fitting the Michaelis-Menten equation to initial rates measured at several triphosphate concentrations using the program Enzfitter (Biosoft, Cambridge, UK)
-
2, 1 mM DTT and 0.5 mM EGTA, as used for previous kinetic experiments with Klenow fragment and similar template-primer duplexes (ref 1c). Kinetic parameters (25 °C) were obtained by fitting the Michaelis-Menten equation to initial rates measured at several triphosphate concentrations using the program Enzfitter (Biosoft, Cambridge, UK).
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-
-
-
26
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84992238783
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-
(a) Stopped-flow spectrometers from Kin-Tek Instruments (ref 14b), with two identical loading syringes (actuated by either nitrogen gas pressure or a stepping motor) leading to a rapid (˜1.6 ms) mixing chamber connected to a stop syringe, were used. The stop chamber was irradiated by a variable wavelength monochromatic light source set to 340 nm and monitored by a photodiode connected to a computer. Syringe A contained 4 μM Klenow fragment and 20 μM DNA-9-mer/DNA-20-mer (ref 9) in MOPS buffer. Syringe B contained 5-60 μM 4S-TTP in MOPS buffer. Data were collected for time periods ranging from 0.1 s to up to 10 s. The results of several runs at each substrate concentration were averaged
-
(a) Stopped-flow spectrometers from Kin-Tek Instruments (ref 14b), with two identical loading syringes (actuated by either nitrogen gas pressure or a stepping motor) leading to a rapid (˜1.6 ms) mixing chamber connected to a stop syringe, were used. The stop chamber was irradiated by a variable wavelength monochromatic light source set to 340 nm and monitored by a photodiode connected to a computer. Syringe A contained 4 μM Klenow fragment and 20 μM DNA-9-mer/DNA-20-mer (ref 9) in MOPS buffer. Syringe B contained 5-60 μM 4S-TTP in MOPS buffer. Data were collected for time periods ranging from 0.1 s to up to 10 s. The results of several runs at each substrate concentration were averaged.
-
-
-
-
31
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0027493457
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(a)
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(a) Bloom, L. B.; Otto, M. R.; Beechem, J. M.; Goodman, M. F. Biochemistry 1993, 32, 11247.
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Bloom, L.B.1
Otto, M.R.2
Beechem, J.M.3
Goodman, M.F.4
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32
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0029085240
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(b)
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(b) Frey, M. W.; Sowers, L. C.; Millar, D. P.; Benkovic, S. J. Biochemistry 1995, 34, 9185.
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, vol.34
, pp. 9185
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Frey, M.W.1
Sowers, L.C.2
Millar, D.P.3
Benkovic, S.J.4
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