Engineering Vibrio fischeri transcriptional activator LuxR for diverse transcriptional activities.
OBJECTIVE
To alter DNA binding specificity of Vibrio fischeri LuxR and to expand the toolbox for constructing synthetic networks.


RESULTS
A mutation library (about 10,000 individuals) of the DNA binding domain of LuxR were generated. A genetic selection was performed to obtain LuxR mutants that recognize three lux box DNA variants that are not recognized by wild-type LuxR. Six LuxR mutants were identified. The evolved LuxR mutants were further characterized by measuring the transcriptional activities of different combinations of LuxR mutants and lux box variants. Varied transcriptional activities were found in these LuxR-lux box pairs. The background expressions of the evolved LuxR-lux box systems are more tightly regulated than the wild-type LuxR-lux box system.


CONCLUSIONS
The LuxR transcriptional system was evolved to recognize three lux box DNAs which are not recognized by wild-type LuxR.
School of Life Sciences, University of Science and Technology of China, Hefei, 230027, China. ly08207@mail.ustc.edu.cn.
Yang
Y
Lu
D048908:Aliivibrio fischeri / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics / Q000502:physiology; D058615:Synthetic Biology / Q000379:methods; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation / Q000235:genetics / Q000502:physiology
D015534:Trans-Activators
Directed evolution; Genetic selection; Lux box; LuxR; Synthetic biology; Transcriptional activator