Compositions of aminoacyl-tRNA synthetase and uses thereof

• US 7,883,866 B2
• Filed: 03/15/2010
• Issued: 02/08/2011
• 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
• Primary Examiner(s): Rao; Manjunath N
• Assistant Examiner(s): Gebreyesus; Kagnew H
• Application Number:
  US12/724,254
• Publication Number:
  US 20100173379A1
• Time in Patent Office: 330 Days
• Field of Search: None
• US Class Current: 435/69.1
• 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 cell-free composition comprising a nucleic acid sequence encoding the aminoacyl tRNA synthetase (RS) of SEQ ID NO:

5, and the complementary polynucleotide sequence thereof.

2. The composition of claim 1, further comprising a tRNA, wherein said tRNA is preferentially aminoacylated by the RS.

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. The composition of claim 2, wherein said tRNA is enzymatically aminoacylated by the RS.

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

7. The composition of claim 6, wherein said amino acid is a non- naturally encoded amino acid.

8. The composition of claim 7, wherein said non-naturally encoded amino acid is para-acetylphenylalanine.

9. The composition of claim 6, wherein the tRNA is derived from an archael tRNA.

10. The composition of claim 6, wherein the tRNA is derived from M. janneschii.

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

12. The composition of claim 11, wherein said translation system is a cell lysate.

13. The composition of claim 11, wherein said translation system is a reconstituted system.

14. The composition of claim 11, wherein said translation system is a cellular translation system.