In situ mutagenesis and chemotactic selection of microorganisms in a diffusion gradient chamber.
A new method has been developed to rapidly generate and select microbial strains having increased resistance to an inhibitory compound. The method combines in situ mutagenesis with use of a continuous gradient of the inhibitor to sort cells according to their resistance levels. Microbial chemotaxis is induced to accelerate the selection process. The method was used to develop a strain of E. coli having a feedback-resistant DAHP synthase enzyme. An unsteady-state mathematical model of the process has been developed. The model, that can reproduce key trends observed experimentally, was used to explore the effects of chemotaxis on the efficiency of the selection process.
Mikola
M R
MR
Department of Chemical Engineering, Michigan State University, East Lansing 48824, USA.
Widman
M T
MT
Worden
R M
RM
D010723:3-Deoxy-7-Phosphoheptulonate Synthase / Q000032:analysis*; D002633:Chemotaxis; D015194:Diffusion Chambers, Culture; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000528:radiation effects; D005947:Glucose / Q000737:chemistry; D008962:Models, Theoretical; D016297:Mutagenesis, Site-Directed; D014466:Ultraviolet Rays
D010723:3-Deoxy-7-Phosphoheptulonate Synthase; D005947:Glucose