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Fig. 4 | Microbial Cell Factories

Fig. 4

From: Identification of a novel family B DNA polymerase from Enterococcus phage IME199 and its overproduction in Escherichia coli BL21(DE3)

Fig. 4

Biochemical characteristics of IME199 DNAP. A Effect of different metal ions on the polymerase activity of IME199 DNAP. 400 nM primed-template (24/50 nt) substrate was incubated with 50 nM IME199 DNAP and 125 μM dNTPs at 30 °C for 20 min in reaction buffers containing different metal ions (final concentration of 10 mM), and then analyzed on a 20% denaturing PAGE gel. B Effect of temperature on the polymerase activity of IME199 DNAP. 400 nM primed-template (24/50 nt) substrate was incubated with 50 nM IME199 DNAP and 125 μM dNTPs at different temperatures for 20 min in reaction buffers containing Mg2+, and then analyzed on a 20% denaturing PAGE gel. C Effect of different pH values on the polymerase activity of IME199 DNAP. 400 nM primed-template (24/50 nt) substrate was incubated with 50 nM IME199 DNAP and 125 μM dNTPs at 30 °C for 20 min in different pH reaction buffers, and then analyzed on a 20% denaturing PAGE gel. The primed-template (24/50 nt) substrate as same as Fig. 3

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Microbial Cell Factories

ISSN: 1475-2859

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