Abstract
The cloning and expression of toxic proteins in bacteria have posed a great challenge because of the leaky expression in inducible expression systems. Using artificial gene synthesis and clone screening methods, we identified a mutant T5 promoter, which significantly reduced leaky expression of lac operator. The mutant T5 promoter contains two T deletions at −35 region and may reduce promoter activity. A bacterial lethal gene, Φ174 lytic gene E, was successfully cloned in this system and expressed in the presence of isopropyl β-d-1-thiogalactopyranoside. The system is compatible with existing T5 inducible expression systems and can be used for the controlled expression of toxic proteins.
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Acknowledgments
This work was supported by Grant 30500563 from the National Science Foundation of China, Grant NCET-06-0615 (NECT), Grant “A Foundation for the Author of National Excellent Doctoral Dissertation of PR China” from the Ministry of Education of the Republic of China.
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Guo, J., Jia, R. A novel inducible expression system for the functional study of toxic gene in bacteria. World J Microbiol Biotechnol 30, 1527–1531 (2014). https://doi.org/10.1007/s11274-013-1573-0
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DOI: https://doi.org/10.1007/s11274-013-1573-0