Development of a novel uric-acid-responsive regulatory system in Escherichia coli
- 522 Downloads
A novel uric-acid-responsive regulatory system was developed in Escherichia coli by adapting the HucR-related regulatory elements from Deinococcus radiodurans into E. coli. The induction performance of this system was compared to the performance of both the pBAD and pET systems. Our novel regulatory system was induced in a dose-dependent manner in the presence of uric acid and exhibited low basal expression in its absence. The system was characterized by a wide dynamic range of induction, being compatible with various E. coli strains and not requiring genomic modifications of the bacterial host. E. coli DH5α and DH10B were the most suitable host strains for optimal performance of this system. In conclusion, we developed a regulatory system with potential for applications in both recombinant protein expression and metabolic optimization.
KeywordsRegulatory protein Uric acid Dynamic range Induction fold Escherichia coli
The authors would like to thank Mr. Ye Mao for his technical assistance in this research. This work was supported by the Ministry of Science and Technology of China Grant 2013CB734003, the National Natural Science Foundation of China Grant 21172095, and the Key Research Program of the Chinese Academy of Sciences Grant KSZD-EW-Z-015-2.
- Karin H, Ruhdal JP (1998) Artificial promoter libraries for selected organisms and promoters derived from such libraries. Patent No. WO1998007846Google Scholar
- Siegele DA, Hu JC (1997) Gene expression from plasmids containing the araBAD promoter at subsaturating inducer concentrations represents mixed populations. Proc Natl Acad Sci U S A 94:8168–8172Google Scholar
- Tabor S (2001) Expression using the T7 RNA polymerase/promoter system. In: Ausubel FA, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) Current protocols in molecular biology. Wiley, New York, pp 16.2.1–16.2.11Google Scholar