FIG 1.

Biochemical analyses of G-N-7 MTase activity of PEDV nsp14 in vitro. (A) SDS-PAGE analysis of purified PEDV nsp14 protein. The 6×His-tagged PEDV nsp14 was expressed and purified from E. coli. (B) Analysis of G-N-7 MTase activity of PEDV nsp14 in different pHs. One microgram of purified PEDV nsp14 was incubated with 2 × 10³ cpm of Gp*ppA-RNA₄₃ substrate in methylation buffer from pH 4.0 to 10.0 (lanes 3 to 9) at 37°C for 4 h. The products were digested with nuclease P1 at 50°C for 30 min and separated on a TLC plate. The migration of the markers m⁷Gp*ppA and Gp*ppA (lanes 1 and 2) is indicated. Spots were dried and visualized with a phosphorimager.