Expression, purification, and characterization of SARS coronavirus RNA polymerase

摘要: The RNA-dependent RNA polymerase (RdRp) of SARS coronavirus (SARS-CoV) is essential for viral replication and a potential target anti-SARS drugs. We report here the cloning, expression, purification N-terminal GST-fused SARS-CoV RdRp its catalytic domain in Escherichia coli. During purification, full-length GST-RdRp was found to cleave into three main fragments: an p12 fragment, middle p30 C-terminal p64 fragment comprising domain, presumably due bacterial proteases. Biochemical assays show that has activity fragments form complex exhibits comparable activity, whereas GST-p64 protein no suggesting required possibly via involvement template-primer binding. Nonnucleoside HIV-1 RT inhibitors are shown have evident inhibitory effect on activity. This work provides basis biochemical structural studies development