Isolation, characterization and molecular cloning of Duplex-Specific Nuclease from the hepatopancreas of the Kamchatka crab

Additional file 1:

Additional figure 1. DSN expression in E. coli. SDS-PAGE of protein fractions before and after purification by metal-affinity chromatography. Cont lanes: protein samples obtained from control E. coli strains containing pET; Exp lanes: proteins obtained from E. coli strains expressing pET-DSN. Lane 1 and 2, total protein; lane 3 and 4, protein compositions of inclusion bodies; lane 5 and 6, proteins purified from inclusion bodies by metal-affinity chromatography; lanes 7 and 8, proteins purified by metal-affinity chromatography of soluble fractions. M denotes the markers. Molecular masses of standards are indicated at left.

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Additional file 2:

Additional figure 2. Elution profile of DSN on Heparin Sepharose. The heparin Sepharose column (1.5 × 20 cm) was equilibrated with 50 mM Tris-HCl, pH 7.15. Proteins were eluted using stepwise increases in NaCl concentration (50 mM, 100 mM and 1 M) at a flow rate of 1.5 ml/min. Fractions with DNAse activity are indicated in gray.

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Additional file 3:

Additional figure 3. Chromatography of DSN on Sephacryl S-200. A Sephacryl S-200 column (1 × 70 cm) was equilibrated with 50 mM Tris-HCl, pH 7.15 containing 150 mM NaCl and eluted at a flow rate 0.3 ml/min. Fractions with DNAse activity are indicated in grey.

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