In a two-phase operation, E. coli containing λSNNU1 (Q- S -) in the chromosome is typically cultured at 33°C and cloned gene expression is induced by elevating the temperature. At least 40°C is necessary for complete induction of cloned gene expression; however, temperatures above 40°C have been shown to inhibit cloned gene expression. This suggests that a three-phase operation, which has an induction phase between the growth and production phases, may result in higher gene expression. In this study, optimal temperature management strategies were investigated for the three-phase operation of cloned gene expression in thermally inducible E. coli/bacteriophage systems. The optimal temperature for the induction phase was determined to be 40°C. When the temperature of the production stage was 33°C, the optimal time period for the induction phase at 40°C was determined to be 60 min. In contrast, when the temperature of the production phase was 37°C, the optimal period for the induction phase at 40°C was 20∼30 min. When the three-phase temperature and temporal profile were set at a growth phase of 33°C, an induction phase at 40°C for 30 min, and a production phase at 37°C, the highest level of cloned gene expression was achieved.
Bibliographical noteFunding Information:
Acknowledgments This work was supported by Korea Science and Engineering Foundation (R11-2000-075-01004 - 0) through the Nano-Bioelectronics and Systems Research Center, Seoul National University, Seoul, Korea.
- Bacteriophage λ
- Escherichia coli
- Recombinant gene expression
- Temperature induction strategy
- Temperature sensitive mutant