Fig. 2
From: Ultrahigh-throughput screening-assisted in vivo directed evolution for enzyme engineering
Construction and characterization of temperature-controlled in vivo evolution system in E. coli. a Construction of the two plasmids for in vivo mutagenesis. The expression of DNA Pol I* was controlled by a thermal-responsive PR promoter. b Reversion tests were implemented after the two plasmids were cotransformed into E. coli BL21 (DE3). Cm, chloramphenicol; Km, kanamycin; Carb, carbenicillin; Spc, spectinomycin. c Carbenicillin-resistance reversion rate of wild-type strain with/without pol I or pol I* gene. d Reversion rate of cells from the saturated culture at different distances from ColE1 ori