The ompA signal peptide directed secretion of Staphylococcal nuclease A by Escherichia coli.

Use of site-directed mutagenesis to investigate the basis for the specificity of hirudin

“…published elsewhere. The expression vector was constructed essentially as described by Takahara et al (1985). The gene for hirudin was synthesized as previously described (Rink et al, 1984) on the basis of the amino acid sequence (shown in Figure 1) of Dodt et al (1984) and ligated into the pIN-III-ompA2 plasmid (Ghrayeb et al, 1984).…”

Characterization of a soluble, catalytically active form of Escherichia coli leader peptidase: requirement of detergent or phospholipid for optimal activity

“…The cloning of the pro-OmpA nuclease A gene into the IPTG-inducible plasmid pONFl (Takahara et al, 1985) and the overexpression of the protein were described by Chatterjee et al (1995).…”

“…Purification of Pro-OmpA Nuclease A. Pro-OmpA nuclease A, which is a hybrid protein of staphylococcal nuclease A fused to the signal peptide of the OmpA protein (Takahara et al, 1985), was overexpressed using E. coli strain SB221 bearing the plasmid pONFl. The cells were grown to the late log phase in M9-Casamino acids medium supplemented with 5 pg/mL leucine and 5 pg/mL tryptophan.…”

Chemical Optimization of Whole-Cell Transfer Hydrogenation Using Carbonic Anhydrase as Host Protein

“…For the periplasmic compartmentalization, CAII was N-terminally fused to the signal peptide of the outer membrane protein A (OmpA) to ensure its secretion to the periplasm of E. coli (Figure a). , For the surface display, CAII was anchored in the outer membrane by fusing it to a truncated E. coli lipoprotein Lpp (residues 1–9), followed by the first five β-sheets of OmpA (residues 46–159, Figure b). , To probe the localization and functionality of CAII, the fluorescent probe 4 consisting of a carboxyfluorescein linked to an arylsulfonamide (Figure c) was synthesized (see Supporting Information). Probe 4 and a primary anti-CAII antibody in combination with a fluorescent secondary antibody were used to stain E. coli cells containing (i) an empty vector (negative control 1) or plasmids targeting CAII to (ii) the cytoplasm (CAII cytoplasm , negative control 2), (iii) the periplasm (CAII periplasm ), or (iv) the cell surface (CAII surface display ).…”

“…For the periplasmic compartmentalization, CAII was Nterminally fused to the signal peptide of the outer membrane protein A (OmpA) to ensure its secretion to the periplasm of E. coli (Figure 1a). 11,28 For the surface display, CAII was anchored in the outer membrane by fusing it to a truncated E. coli lipoprotein Lpp (residues 1−9), followed by the first five βsheets of OmpA (residues 46−159, Figure 1b). 13,29 To probe the localization and functionality of CAII, the fluorescent probe 4 consisting of a carboxyfluorescein linked to an arylsulfonamide (Figure 1c) was synthesized (see Supporting Information).…”