title: A protocol to express and isolate HRV16 3C protease for use in protease assays.
abstract: Human rhinovirus (HRV) proteases are highly conserved across serotypes and have very similar target specificity. However, there are some serotype-specific differences in their action. It is therefore necessary when performing in vitro protease assays to ensure that the recombinant proteases are specific to the serotype of the HRV under study. We describe a simple method for isolating HRV16 3C protease from a bacterial expression system, including transformation of bacterial cells with a commercially available cDNA plasmid which can be adapted to use for 3C proteases from any other HRV serotypes. The extracted, active 3C protease can then be used for in vitro protease assays.
authors.affiliation: Faculty of Education, Science, Technology and Mathematics, Centre for Research in Therapeutic Solutions, University of Canberra, 1 Kirinari Street, Bruce, Canberra, ACT, 2601, Australia.
authors.forename: Lora M
authors.initials: LM
authors.lastname: Jensen
authors.forename: Erin J
authors.initials: EJ
authors.lastname: Walker
authors.forename: Reena
authors.initials: R
authors.lastname: Ghildyal
mesh_terms: D000086042:3C Viral Proteases; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D006367:HeLa Cells / Q000821:virology; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids; D014169:Transformation, Bacterial; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D014764:Viral Proteins; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases