Genetic element that enhances protein translation

Patent metadata

Field	Value
Publication number	US-9234197-B2
Application number	US-201214349835-A
Country	US
Kind code	B2
Filing date	Oct 26, 2012
Priority date	Oct 28, 2011
Publication date	Jan 12, 2016
Grant date	Jan 12, 2016

Abstract

The present invention provides isolated polynucleotides that can serve as translation enhancing elements and their use in protein expression reagents and methods.

First claim

We claim:

1. A recombinant viral or plasmid expression vector comprising
(a) a translation enhancement element (TEE) of between 37 and 42 nucleotides in length, comprising the nucleic acid sequence of 5′-CATATTGAAGAGACAGAGTGATATATAAAACTGCTAA-3′ (SEQ ID NO: 1); and
(b) a cloning site located downstream of the TEE.

2. The expression vector of claim 1 , further comprising a promoter located upstream of the TEE.

3. The expression vector of claim 1 , further comprising a protein encoding nucleic acid cloned into the cloning site.

4. An isolated recombinant host cell comprising the expression vector of claim 1 .

5. A method for protein expression, comprising contacting the expression vector of claim 3 with reagents and under conditions for promoting expression of the protein encoded by the protein-encoding nucleic acid, and expressing the protein.

6. The method of claim 5 , wherein the protein is expressed in a cell-free translation system.

7. The method of claim 5 , wherein the protein is expressed by a recombinant host cell.

8. The method of claim 5 , wherein the expression vector is a recombinant vaccinia virus.

9. The method of claim 7 , wherein the recombinant host cell expresses the protein using non-viral-mediated translation.

10. An isolated recombinant virus, comprising the expression vector of claim 1 , wherein the expression vector is comprises a viral expression vector.

11. The recombinant virus of claim 10 , wherein the recombinant virus is a vaccinia virus, and the viral expression vector is a vaccinia virus expression vector.

12. The expression vector of claim 1 , wherein the TEE consists of 5′-CATATTGAAGAGACAGAGTGATATATAAAACTGCTAA-3′ (SEQ ID NO: 1).

13. The expression vector of claim 1 , wherein the TEE consists of 5′-AGAACCATATTGAAGAGACAGAGTGATATATAAAACTGCTAA-3′ (SEQ ID NO:2).

Assignees

• Univ Arizona
• Univ Arizona

Inventors

• Chaput John
• Jacobs Bertram
• Wellensiek Brian
• Flores Julia

Classifications

• C12N2830/30

  being an enhancer not forming part of the promoter region · CPC title

• C12N15/67

  General methods for enhancing the expression · CPC title

• C12N15/85 Primary

  for animal cells · CPC title

• C12N2840/105

  enhancing translation · CPC title

• C12N2830/85

  mammalian · CPC title