FUNCTIONAL BIOMOLECULES AND METHODS

• US 20120165225A1
• Filed: 05/15/2008
• Published: 06/28/2012
• Est. Priority Date: 05/15/2007
• Status: Active Grant

• Current Assignee: BioTex Incorporated
• Original Assignee: BioTex Incorporated
• Inventors: Stepanov, Victor G., Fox, George E., Jackson, George W., McNichols, Roger J., Liu, Yamei
• Granted Patent: US 8,916,376 B2
• Time in Patent Office: Days
• US Class Current: 506/14
• CPC Class Codes: C12N 15/1048 SELEX

Abstract

The present invention is generally directed to novel functionalized biomolecules and methods for generating such biomolecules. Biomolecules may generally include nucleic acids, peptides, multicomponent molecular complexes and/or any other molecular products that may be produced by living organisms. The present invention is further directed to cells and/or organisms manipulated to produce such functionalized biomolecules. The cells contemplated by the present invention include both prokaryotic as well as eukaryotic cells. The functionalized biomolecules are produced via materials introduced into the cell using standard molecular biology techniques or are incorporated within the genomic nucleic acid of a cell by standard recombination techniques. Further contemplated is the use of such cells for sequestration of target molecules within the cells.

Claims

1. An expression vector comprising:

a chimeric gene encoding a peptide, operatively linked to a functional promoter, said expression vector when inserted into a host transcribes said chimeric gene into a gene product which is translated into said peptide which is capable of binding to or altering at least one target molecule which induces stress on said host;

wherein said peptide comprises a non-naturally occurring, randomly selected peptide which is not derived from a naturally occurring peptide and is selected by sequential culturing of a host exposed to said target molecule and identifying conferred resistance to said stress on said host caused by said target molecule.

2. The expression vector of claim 1, wherein said vector further comprises at least one of a selection marker or a marker for selective induction.

3. The expression vector of claim 2, wherein said vector further comprises at least one selection marker and wherein said selection marker is an antibiotic resistance marker.

4. The expression vector of claim 1, wherein said promoter is a T7 RNA polymerase or a ribosomal RNA promoter.

5. The expression vector of claim 1, wherein said biomolecule ligand includes an amino acid sequence selected from the group of SEQ ID NO 1, MSHAYFVCNRCDSSNHSAHE or SEQ ID NO 2, MSHATATPASRRRLPLRS.

6. The expression vector of claim 1, wherein said at least one target molecule is a waste fluid contaminant.

7. The expression vector of claim 6, wherein said waste fluid contaminant is at least one of an inorganic molecule, an organic molecule, a toxin, a peptide, or a viral particle.

8. The expression vector of claim 1, wherein said at least one target molecule is at least one of hormones, antibodies, proteins, enzymes, pharmaceuticals or metals.

9. An isolated cell comprising said expression vector of claim 1.

10. The cell of claim 9, wherein said cell is a prokaryotic cell or a eukaryotic cell.

11. An isolated cell comprising:

at least one artificially inserted nucleic acid sequence encoding mRNA, said mRNA encoding a peptide which binds to or catalytically alters a target molecule which induces stress on said cell;

wherein said at least one nucleic acid sequence is selected by sequential culturing of cells containing said sequence with exposure to at least one target molecule and identifying conferred to resistance to said stress caused by said target molecule.

12. The cell of claim 11, wherein said peptide includes an amino acid sequence selected from the group of SEQ ID NO 1, MSHAYFVCNRCDSSNHSAHE or SEQ ID NO 2, MSHATATPASRRRLPLRS.

13. The cell of claim 11, wherein said cell is a prokaryotic cell or a eukaryotic cell.

14. The cell of claim 11, wherein said target molecule is a waste fluid contaminant.

15. The cell of claim 14, wherein said waste fluid contaminant is an inorganic molecule, an organic molecule, a toxin, a peptide, or a viral particle.

16. The cell of claim 11, wherein said target molecule is a hormone, an antibody, a protein, an enzyme, or a metal.

17-20. -20. (canceled)

21. A set of isolated cells comprising:

a plurality of cells, each of said cells comprising at least one artificially inserted nucleic acid sequence encoding mRNA;

wherein each of said plurality of cells comprises a different artificially inserted nucleic acid sequence encoding mRNA, at least one of which encodes a non-naturally derived peptide which binds to a desired target molecule and confers an identifiable resistance to stress induced by said desired target molecule to one of said plurality of cells.

22. The set of isolated cells of claim 21, wherein said non-naturally derived peptide comprises a randomly selected amino acid sequence.

23. The set of isolated cells of claim 21, wherein said non-naturally derived peptide comprises a non-antibody peptide sequence.

24. The set of isolated cells of claim 21, wherein said resistance to stress caused by said desired target molecule comprises survivability of at least one of said plurality of cells when exposed to said desired target molecule.