Claims
- 1. A method of affixing a peptide acceptor to an RNA molecule, said method comprising:
(a) providing an RNA molecule having a 3′ sequence which forms a hairpin structure; (b) providing a peptide acceptor covalently bonded to a nucleic acid linker molecule; and (c) hybridizing said RNA molecule to said nucleic acid linker molecule under conditions which allow covalent bond formation to occur between said peptide acceptor and said RNA molecule.
- 2. The method of claim 1, wherein said covalent bond formation is catalyzed by T4 DNA ligase.
- 3. A method of affixing a peptide acceptor to an RNA molecule, comprising:
(a) providing an RNA molecule; (b) providing a peptide acceptor covalently bonded to a nucleic acid linker molecule, wherein the 5′ sequence of said linker molecule forms a hairpin structure; and (c) hybridizing said RNA molecule to said nucleic acid linker molecule under conditions which allow covalent bond formation to occur between said peptide acceptor and said RNA molecule.
- 4. The method of claim 3, wherein said covalent bond formation is catalyzed by T4 DNA ligase.
- 5. The method of claim 1 or 3, wherein said RNA molecule comprises a translation initiation sequence and a start codon operably linked to a protein coding sequence.
- 6. The method of claim 1 or 3, wherein said peptide acceptor is puromycin.
- 7. The method of claim 1 or 3, wherein said peptide acceptor is covalently bonded to a non-nucleotide linker molecule.
- 8. The method of claim 1 or 3, wherein said linker comprises triethylene glycol spacers.
- 9. The method of claim 1 or 3, wherein said linker comprises 2′-OMe-RNA phosphoramidites.
- 10. The method of claim 1 or 3, wherein said RNA or said linker contains an affinity purification sequence and said method further comprises purifying said RNA.
- 11. The method of claim 10, wherein said affinity purification sequence comprises a poly (A) sequence.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. Ser. No. 09/619,103, filed Jul. 19, 2000, which claims the benefit of the filing date of provisional application, U.S. Ser. No. 60/145,834, filed Jul. 27, 1999, now abandoned, all hereby incorporated by reference.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60145834 |
Jul 1999 |
US |
Divisions (1)
|
Number |
Date |
Country |
Parent |
09619103 |
Jul 2000 |
US |
Child |
10208357 |
Jul 2002 |
US |