Ultrahigh-throughput screening-assisted in vivo directed evolution for enzyme engineering.

ABSTRACT:

Background

Classical directed evolution is a powerful approach for engineering biomolecules with improved or novel functions. However, it traditionally relies on labour- and time-intensive iterative cycles, due in part to the need for multiple molecular biology steps, including DNA transformation, and limited screening throughput.

Results

In this study, we present an ultrahigh throughput in vivo continuous directed evolution system with thermosensitive inducible tunability, which is based on error-prone DNA polymerase expression modulated by engineered thermal-responsive repressor cI857, and genomic MutS mutant with temperature-sensitive defect for fixation of mutations in Escherichia coli. We demonstrated the success of the in vivo evolution platform with β-lactamase as a m

SUBMITTER: Chen S 

PROVIDER: S-EPMC10804518 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature