Expression, purification, and characterization of SARS coronavirus RNA polymerase

Purification and Characterization of RNA Polymerase from

We have purified and characterized a DNA-dependent RNA polymerase from the blue-green alga Fremyela dipsiphon. This enzyme, purified by gel filtration, DEAE-ceflulose chromatography, and glycerol gradient centrifugation, is comprised of five polypeptide subunits. Their masses are 161,000, 134,000, 91,000, 72,000, and 41,000 daltons. Preparative electrophoresis of the purified enzyme on nondenat...

Ultrastructural Characterization of SARS Coronavirus

Severe acute respiratory syndrome (SARS) was first described during a 2002-2003 global outbreak of severe pneumonia associated with human deaths and person-to-person disease transmission. The etiologic agent was initially identified as a coronavirus by thin-section electron microscopic examination of a virus isolate. Virions were spherical, 78 nm in mean diameter, and composed of a helical nucl...

SARS diagnosis, monitoring and prognostication by SARS-coronavirus RNA detection.

A second, non-canonical RNA-dependent RNA polymerase in SARS coronavirus.

In (+) RNA coronaviruses, replication and transcription of the giant approximately 30 kb genome to produce genome- and subgenome-size RNAs of both polarities are mediated by a cognate membrane-bound enzymatic complex. Its RNA-dependent RNA polymerase (RdRp) activity appears to be supplied by non-structural protein 12 (nsp12) that includes an RdRp domain conserved in all RNA viruses. Using SARS ...

Purification and Characterization of RNA Polymerase from Fremyella diplosiphon.

We have purified and characterized a DNA-dependent RNA polymerase from the blue-green alga Fremyella diplosiphon. This enzyme, purified by gel filtration, DEAE-cellulose chromatography, and glycerol gradient centrifugation, is comprised of five polypeptide subunits. Their masses are 161,000, 134,000, 91,000, 72,000, and 41,000 daltons. Preparative electrophoresis of the purified enzyme on nonde...