Abstract
Objectives
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.
Conclusion
The LuxR transcriptional system was evolved to recognize three lux box DNAs which are not recognized by wild-type LuxR.
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Acknowledgments
This work was supported by National High Technology Research and Development Program of China (2012AA02A704) and the Fundamental Research Funds for the Central Universities (Grant No. WK2070000019).
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Lu, Y. Engineering Vibrio fischeri transcriptional activator LuxR for diverse transcriptional activities. Biotechnol Lett 38, 1459–1463 (2016). https://doi.org/10.1007/s10529-016-2134-z
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DOI: https://doi.org/10.1007/s10529-016-2134-z