Optimization of DsRed production in Escherichia coli: Effect of ribosome binding site sequestration on translation efficiency

Abstract DsRed‐Express, a popular reporter protein, cannot be expressed in Escherichia coli using a consensus ribosome binding site (RBS) potentially due to basepairing in the RBS that inhibits translation initiation. Saturation mutagenesis was used to probe for a gene sequence that minimized basepairing in the RBS while maintaining the same spectral properties and maturation characteristics as DsRed‐Express. The new DsRed, designated here as RFP EC for E. coli optimized red fluorescent protein, fluoresces 2.5 times greater than DsRed‐Express when expressed from the same vector. © 2005 Wiley Periodicals, inc.