Hong Kong Med J 2012;18(Suppl 3):S16-9
Impact of SARS-coronavirus-encoded proteins on cellular signalling pathways and cytokine/chemokine gene expression
JKL Chan, P Cheung, M He, Z Wu
Department of Biochemistry, Hong Kong University of Science and Technology, Hong Kong
1. Although GFP-tagged ORF8 and ORF3 potently activate the JNK and p38 MAPK pathways, expression of non-tagged SARS-CoV-encoded ORF8 or ORF3 has no obvious effect.
2. Although both GFP-tagged ORF8 and ORF3 induce cell death, expression of non-tagged ORF8 or ORF3 has no obvious effect on cell survival.
3. Addition of an epitope tag to a protein of interest, a common way to study novel proteins in the absence of suitable antibodies, may generate unexpected artefacts. Caution should be taken with any results derived from epitope-tagged proteins.
4. When studying a novel protein, it is essential to prepare suitable antibodies to facilitate detection and purification (eg by immunoprecipitation) of the native or endogenous proteins.