Genetic screens and directed evolution for protein solubility.
Overexpressed proteins are often insoluble, and can be recalcitrant to conventional solubilization techniques such as refolding. Directed evolution methods, in which protein diversity libraries are screened for soluble variants, offer an alternative route to obtaining soluble proteins. Recently, several new protein solubility screens have been developed that do not require structural or functional information about the target protein. Soluble protein can be detected in vivo and in vitro by fusion reporter tags. Protein misfolding can be measured in vivo using the bacterial response to protein misfolding. Finally, soluble protein can be monitored by immunological detection. Efficient, well-established strategies for generating and recombining genetic diversity, driven by new screening and selection methods, can furnish correctly folded, soluble protein.
Waldo
Geoffrey S
GS
BN-2, MS M888, Los Alamos National Laboratories, Los Alamos, NM 87545, USA.
D019020:Directed Molecular Evolution / Q000379:methods*; D017930:Genes, Reporter; D023281:Genomics / Q000379:methods; D015335:Molecular Probes; D019151:Peptide Library; D011506:Proteins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D012995:Solubility
D015335:Molecular Probes; D019151:Peptide Library; D011506:Proteins