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Figure 1 | BMC Molecular Biology

Figure 1

From: Biochemical characterization of a recombinant Japanese encephalitis virus RNA-dependent RNA polymerase

Figure 1

Purification of recombinant JEV NS5 protein. JEV NS5 protein was expressed in E. coli and purified by Ni-NTA chromatography, gel filtration chromatography, and ion-exchange chromatography using an SP-Sepharose column. (A) Imidazole elution profile of JEV NS5 from Ni-NTA resin. (B) Gel filtration chromatography elution profile of JEV NS5. (C) NaCl-elution profile of JEV NS5 from an SP-Sepharose column. (A-C) Fractions from each purification step were resolved by SDS-12% PAGE and stained with Coomassie brilliant blue. (D) JEV NS5 and its mutant NS5D668A from a peak fraction eluted from an SP-Sepharose column were resolved by SDS-12% PAGE and visualized by silver staining. (E) Western blot analysis of the purified JEV NS5 and NS5D668A using an anti-His6 antibody. The sizes of protein markers are indicated in kilodaltons. Closed and open arrowheads indicate the full-length JEV NS5, and its major cleaved form identified by MALDI-TOF analysis, respectively.

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