Method of producing a recombinant protein
The present invention relates to a recombinant signal sequence derived from E. coli . The invention also relates to a fusion protein comprising the signal sequence, a recombinant protein and methods of producing the recombinant protein. The recombinant signal sequence can be used to provide a method for controlling the viscosity of the fermentation, and/or controlling basal (pre-induction) expression of the recombinant protein.
1. A recombinant signal sequence comprising (a) the sequence MGRISSGG (SEQ ID NO: 1) or a variant thereof that differs in sequence by 1 amino acid substitution, deletion or insertion, and (b) a heterologous signal sequence immediately C-terminal to (a).
2. A recombinant fusion protein comprising (a) the recombinant signal sequence of claim 1 ; and (b) a heterologous recombinant protein C-terminal to (a).
3. The recombinant signal sequence of claim 1 , wherein the signal sequence comprises MGRISSGG (SEQ ID NO: 1).
4. The recombinant signal sequence of claim 1 , wherein the signal sequence comprises the variant of MGRISSGG (SEQ ID NO: 1) that differs in sequence by 1 amino acid substitution, deletion or insertion.
5. The recombinant signal sequence of claim 1 , wherein the heterologous signal sequence is an Escherichia coli signal sequence.
6. The recombinant signal sequence of claim 5 , wherein the heterologous signal sequence is a periplasmic signal sequence from Escherichia coli.
7. The recombinant signal sequence of claim 5 , wherein the heterologous signal sequence is selected from the group consisting of a OmpA signal sequence, a MalE signal sequence, a PelB signal sequence, a OmpT signal sequence, and a LamB signal sequence.
8. The recombinant signal sequence of claim 1 , wherein the heterologous signal sequence comprises a sequence selected from the group consisting of SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, and SEQ ID NO: 40.
9. The recombinant signal sequence of claim 8 , wherein the heterologous signal sequence comprises SEQ ID NO: 33.
10. The recombinant signal sequence of claim 9 , wherein the heterologous signal sequence consists of SEQ ID NO: 33.
11. The recombinant signal sequence of claim 1 , wherein the recombinant signal sequence comprises the sequence MGRISSGGMKKTAIAIAVALAGFATVAQA (SEQ ID NO: 55).
12. The recombinant signal sequence of claim 1 , wherein the recombinant signal sequence consists of the sequence MGRISSGGMKKTAIAIAVALAGFATVAQA (SEQ ID NO: 55).
13. The recombinant fusion protein of claim 2 , wherein the recombinant signal sequence comprises the sequence MGRISSGGMKKTAIAIAVALAGFATVAQA (SEQ ID NO: 55).
14. The recombinant fusion protein of claim 2 , wherein the recombinant signal sequence consists of the sequence MGRISSGGMKKTAIAIAVALAGFATVAQA (SEQ ID NO: 55).
15. The recombinant fusion protein of claim 2 , wherein the heterologous recombinant protein comprises an antigen binding protein.
16. The recombinant fusion protein of claim 15 , wherein the antigen binding protein is selected from the group consisting of a monoclonal antibody, an antibody fragment, and a domain antibody (dAb).
17. The recombinant fusion protein of claim 2 , wherein the heterologous recombinant protein comprises a viral protein.
18. The recombinant fusion protein of claim 2 , wherein the heterologous recombinant protein comprises a bacterial toxin or a bacterial toxoid.
19. The recombinant fusion protein of claim 2 , wherein the heterologous recombinant protein comprises a cancer antigen.