A lot of the work we do in the laboratory is trying to find better ways to do Directed Evolution. One of the thing we believe is that expressing libraries in their native genomic context let you navigate the fitness landscape better.
If you want to find out how exactly we build those libraries, the detailed protocols we use, both in E. coli and in S. thermophilus have now been published as a book chapter by Springer Protocols.
“Dorrazehi G.M., Worms S., Chirakadavil J.B., Mignolet J., Hols P., Soumillion P. (2020) Building Scarless Gene Libraries in the Chromosome of Bacteria. In: Iranzo O., Roque A. (eds) Peptide and Protein Engineering. Springer Protocols Handbooks. Humana, New York, NY
Open access:
https://dial.uclouvain.be/pr/boreal/object/boreal:231618
Chromosomal Libraries Chapter is out!