Dual inducible system for the condensed single protein production system
The present invention provides a dual inducible system for single protein production, as well as a method of inducing high level protein expression using amino acids.
1. A system for expressing a single target protein in an E. coli cell comprising:
(i) an isolated E. coli tryptophan auxotroph; and
(ii) an expression vector comprising an isolated nucleic acid sequence encoding E. coli MazF protein with W14F and W83L substitutions, a sequence-specific mRNA endoribonuclease.
2. The system of claim 1 , further comprising an expression vector comprising an isolated nucleic acid sequence encoding a target protein.
3. The system of claim 1 , further comprising a composition suitable to induce the sequence-specific mRNA endoribonuclease.
4. The system of claim 2 further comprising a composition comprising tryptophan.
5. A system for expressing a single target protein in an E. coli cell comprising:
(i) an isolated E. coli histidine auxotroph; and
(ii) an expression vector comprising an isolated nucleic acid sequence encoding E. coli MazF protein with G27K and H28R substitutions, a sequence-specific mRNA endoribonuclease.
6. The system of claim 5 , further comprising an expression vector comprising an isolated nucleic acid sequence encoding a target protein.
7. The system of claim 5 , further comprising a composition suitable to induce the sequence-specific mRNA endoribonuclease.
8. The system of claim 7 , further comprising a composition comprising histidine.