Prokaryotic expression and solubilisation of Arabidopsis ROOT UVB SENSITIVE 1 from inclusion bodies in Escherichia coli

The RUS (ROOT UVB SENSITIVE 1) proteins characterized by their unique DUF647 domain are widely distributed in eukaryotes. Their functional roles largely unknown except for the possible involvement of Arabidopsis RUS1 and RUS2 early seedling development. To investigate biochemical proteins, full length truncated were seamlessly fused with GFP cloned into prokaryotic expression vector pQE-100 which allows expressed an N-terminal 6?His tag. Expression RUS1-GFP could not be detected after adding inducer IPTG, while a was at high levels formed inclusion bodies Escherichia coli. dissolved denaturing buffer, then fusion protein supernatant bound to Ni-NTA slurry. eluted non-specific binding washed away. purified as single clear band expected size SDS-PAGE, further confirmed Western blot test. Our results suggest that impossible E. coli can form, effectively solubilized.