Article
Directed evolution of copper nitrite reductase to a chromogenic reductant.
Protein engineering, design & selection : PEDS - 1 Mar 2010
MacPherson Iain S, Rosell Federico I, Scofield Melanie, Mauk A Grant, Murphy Michael E P
Abstract excerpt
Directed evolution methods were developed for Cu-containing nitrite reductase (NiR) from Alcaligenes faecalis S-6. The PCR cloning strategy allows for the efficient production of libraries of 100 000 clones by a modification of a megaprimer-based whole-plasmid synthesis reaction. The high-throughput screen includes colony lift onto a nylon membrane and subsequent lysis of NiR-expressing colonies in the presence...
Topics
- Alcaligenes faecalis
- Azurin
- Chromogenic Compounds
- Copper
- Crystallography, X-Ray
- Dianisidine
- Directed Molecular Evolution
- Electrochemistry
- Electrons
- Enzyme Assays
- High-Throughput Screening Assays
- Models, Molecular
- Mutagenesis, Site-Directed