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. 2004 Aug 10;101(34):12694–12699. doi: 10.1073/pnas.0403127101

Fig. 6.

Fig. 6.

Activities of mutant forms of human coronavirus NendoU. (A) Coomassie brilliant blue-stained SDS-polyacrylamide gel showing the expression and purification of HCoV-229E (HCoV) and SARS-CoV (SCoV) MBP-nsp15 fusion proteins. Lanes 1 and 2, total lysates of E. coli cells transformed with pMal-HCoV-nsp15 plasmid DNA and grown in the absence (lane 1) or presence (lane 2) of isopropyl β-d-thiogalactoside (IPTG); lanes 3-12, purified MBP-nsp15 fusion proteins with nsp15 wild-type sequence (WT) or with the indicated substitutions of conserved residues. Numbering of residues is according to their positions in the replicase polyproteins of HCoV-229E and SARS-CoV, respectively. (B) HCoV-229E and SARS-CoV MBP-nsp15 fusion proteins or their mutant derivatives were incubated with dsRNA2, and reaction products were separated in a 20% polyacrylamide gel containing 7.5 M urea.