Sampling the cultivation parameter space for the bacterial production of TLR1 intracellular domain reveals the multiple optima.
T7 expression system is an extremely popular approach for the recombinant protein production in Escherichia coli for structural and functional studies and therapeutic applications. There are many useful tools and successful techniques that allow expressing the desired protein in this system. However, high yield of soluble protein often requires a systematic optimization of a wide range of cell cultivation parameters. Here we analyze the effect of three key cultivation parameters - chemical inductor, temperature and time of post-induction culturing on the expression level of TLR1 intracellular TIR domain in a soluble form. In addition, the influence of Triton X-100 detergent on the protein solubility during the cell lysis was investigated. We show that a high expression level of the correctly folded soluble protein can be obtained under different combinations of cultivation parameters.
Список научных проектов, где отмечена публикация
- Elucidating the structural basis of toll-like receptor signaling by NMR in solution (January 6, 2020 December 31, 2021). . Grant, RFBR.