Synthetic transcription factor and uses thereof
The present invention provides for a synthetic transcription factor (TF) comprising a first peptide capable of binding a target ligand, a second peptide capable of binding a target DNA, and a peptide linker linking the first and second peptides. The present invention also provide for a system for modulating the mutagenesis frequency of a host cell. The host cell has a mutator rate (R) which is inversely proportional to a phenotypic trait (P).
1. A system for modulating mutagenesis frequency of a host cell, comprising a host cell comprising: (a) a synthetic transcription factor (TF) comprising a first peptide capable of binding a target ligand, a second peptide capable of binding a target DNA, and a peptide linker linking the first and second peptides, wherein the first peptide and the second peptide are from different proteins, and (b) a mutator module comprising a target promoter operably linked to a gene that is capable of increasing a mutation rate of the host cell; wherein the host cell has a mutator rate (R) which is inversely proportional to a phenotypic trait (P); wherein the first peptide is a ligand-binding domain of Idi, Idi1 or Erg20, and P is isoprenoid or lycopene production, and (i) the second peptide is a DNA-binding domain (DBD) of AraC and the target promoter is an araBAD promoter and the host cell is a prokaryotic cell, or (ii) the second peptide is a DNA-binding domain (DBD) of Gal 4 and the target promoter is a GAL promoter and the host cell is a yeast cell.
2. The system of claim 1 wherein the host cell comprises a nucleic acid comprising a nucleotide sequence encoding the synthetic TF.
3. The system of claim 2 wherein the host cell comprises a vector capable of stable maintenance of the nucleic acid.
4. The system of claim 3 wherein the vector is an expression vector.
5. The system of claim 1 , wherein (i) when the host cell is a prokaryotic cell the gene that is capable of increasing a mutation rate of the host cell is Escherichia coli mutD 5 , Bacillus subtilis mutM, Pseudomona aeruginosa mutS, Pseudomonas aeruginosa mutL, or Synechococcus sp. mutS, and (ii) when the host cell is a yeast cell the gene that is capable of increasing a mutation rate of the host cell is Saccharomyces cerevisiae msh2, or Saccharomyces cerevisiae him1.
6. The system of claim 1 , wherein the first peptide is a ligand-binding domain of Idi and the second peptide is a DNA-binding domain (DBD) of AraC and the target promoter is the araBAD promoter or the second peptide is a DNA-binding domain (DBD) of Gal4 and the target promoter is a GAL promoter.
7. The system of claim 6 , wherein the GAL promoter is P GAL10 .
8. The system of claim 1 , wherein the first peptide is a ligand-binding domain of Idi, Idi1 or Erg20, and the second peptide is a DNA-binding domain (DBD) of Gal4 and the target promoter is a GAL promoter.
9. The system of claim 8 , wherein the GAL promoter is P GAL10 .
10. The system of claim 1 , wherein the host cell is a prokaryotic cell which is a species of Escherichia, Enterobacter, Azotobacter, Erwinia, Bacillus, Pseudomonas, Klebsiella, Proteus, Salmonella, Serratia, Shigella, Rhizobia, Vitreoscilla , or Paracoccus.
11. The system of claim 10 , wherein the prokaryotic cell is a species of Escherichia.
12. The system of claim 11 , wherein the prokaryotic cell is Escherichia coli.
13. The system of claim 1 , wherein the host cell is a yeast cell which is a species of Saccharomyces or Candida.
14. The system of claim 13 , wherein the yeast cell is a species of Saccharomyces.
15. The system of claim 14 , wherein the yeast cell is Saccharomyces cerevisiae.