Compositions of Aminoacyl-tRNA Synthetase and Uses Thereof

• US 20100173380A1
• Filed: 03/15/2010
• Published: 07/08/2010
• Est. Priority Date: 12/22/2004
• Status: Active Grant

• Current Assignee: Ambrx, Inc. (Johnson & Johnson)
• Original Assignee: Ambrx, Inc. (Johnson & Johnson)
• Inventors: PAULSEL, Andrew, CHO, Ho Sung
• Granted Patent: US 7,838,265 B2
• Time in Patent Office: Days
• US Class Current: 435/183
• CPC Class Codes: C12N 9/93 Ligases (6)
• CPC Class Codes: C12P 21/02 having a known sequence of two or more amino acids, e.g. glutathione

Abstract

Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNA'"'"'s, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNA'"'"'s/synthetases are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins using these orthogonal pairs.

Claims

1. A composition comprising an aminoacyl tRNA synthetase (RS) nucleic acid sequence selected from the group consisting of a polynucleotide sequence encoding the amino acid sequence of SEQ ID NO:

17, and a complementary polynucleotide sequence thereof.

2. The composition of claim 1, wherein said RS preferentially aminoacylates a tRNA.

3. The composition of claim 2, wherein said tRNA is aminoacylated with a non-naturally encoded amino acid.

4. The composition of claim 3, wherein said non-naturally encoded amino acid is para-acetylphenylalanine.

5. A composition comprising the RS of claim 1 and a tRNA, wherein said tRNA is chemically aminoacylated.

6. A composition comprising the RS of claim 1 and a tRNA, wherein said tRNA is enzymatically aminoacylated.

7. The composition of claim 6, wherein said tRNA is enzymatically aminoacylated by the RS.

8. The composition of claim 6, wherein said tRNA is enzymatically aminoacylated by a ribozyme.

9. The composition of claim 1, further comprising a tRNA, wherein the tRNA is aminoacylated with an amino acid by the RS.

10. The composition of claim 9, wherein said amino acid is a non-naturally encoded amino acid.

11. The composition of claim 10, wherein said non-naturally encoded amino acid is para-acetylphenylalanine.

12. The composition of claim 9, wherein the tRNA is derived from an archael tRNA.

13. The composition of claim 9, wherein the tRNA is derived from M. janneschii.

14. The composition of claim 1, further comprising a translation system.

15. The composition of claim 14, wherein said translation system is a cell-free translation system.

16. The composition of claim 14, wherein said translation system is a cell lysate.

17. The composition of claim 14, wherein said translation system is a reconstituted system.

18. The composition of claim 14, wherein said translation system is a cellular translation system.

19-45. -45. (canceled)