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US 2003/0228657 A1Published application
Methods for enhancing the translation and expression of recombinant proteins
AssigneeZymoGenetics, Inc.
InventorsTracey A. Pownder, Chung Chan
Life of this patent
Estimating the term…
Abstract
The increased use of nucleotide sequence data mining techniques has amplified the demand for efficient methods of producing recombinant proteins in prokaryotic cells. A strategy is provided for enhancing the synthesis of recombinant amino acid sequences by improving translation from expression cassettes in vitro before producing recombinant hosts.
Claims22
A purified polynucleotide comprising the following nucleic acid sequence:
- A-B-C-D-E
- wherein:
- A is a start codon;
- B is a polynucleotide sequence of 13 nucleic acids, wherein the polynucleotide sequence has homology to SEQ ID NO:2, wherein at least 5 of the 13 nucleic acids are identical to the corresponding position in SEQ ID NO:2, and wherein the nucleic acid sequence does not code for a stop codon;
- C is a polynucleotide sequence selected from the group consisting of:
- a) a polynucleotide sequence having the first 10 nucleic acids of SEQ ID NO:3;
- b) a polynucleotide sequence having the first 13 nucleic acids of SEQ ID NO:3;
- c) a polynucleotide sequence having the first 16 nucleic acids of SEQ ID NO:3;
- and
- d) a polynucleotide sequence having the first 19 nucleic acids of SEQ ID NO:3;
- D is the polynucleotide sequence as shown in SEQ ID NO:4;
- and
- E is a polynucleotide sequence selected from the group consisting of:
- a) SEQ ID NO:5;
- b) SEQ ID NO:6;
- c) SEQ ID NO:7;
- d) SEQ ID NO:8;
- e) SEQ ID NO:9;
- f) SEQ ID NO:10;
- g) SEQ ID NO:11;
- h) SEQ ID NO:
- 12;
- and
- i) SEQ ID NO:13,
- wherein at least 50% nucleotides are either adenine or thymine;
- and
- wherein only E encodes a stop codon.
24 more elements
A purified polynucleotide comprising the following nucleic acid sequence:
- A-B-C-D-E
- wherein:
- A is a start codon;
- B is a polynucleotide sequence of 13 nucleic acids, wherein the polynucleotide sequence has homology to SEQ ID NO:2, wherein at least 5 of the 13 nucleic acids are identical to the corresponding position in SEQ ID NO:2, and wherein the nucleic acid sequence does not code for a stop codon;
- C is a nucleic acid selected from the group consisting of:
- a) adenine;
- b) thymine;
- c) cytosine;
- and
- d) guanine;
- D is the polynucleotide sequence as shown in SEQ ID NO:4;
- and
- E is a polynucleotide sequence selected from the group consisting of:
- a) SEQ ID NO:5;
- b) SEQ ID NO:6;
- c) SEQ ID NO:7;
- d) SEQ ID NO:8;
- e) SEQ ID NO:9;
- f) SEQ ID NO:10;
- g) SEQ ID NO:1;
- h) SEQ ID NO:
- 12;
- and
- i) SEQ ID NO:
- 13,
- wherein at least 50% nucleotides are either adenine or thymine;
- and
- wherein only E encodes a stop codon.
25 more elements
A purified polynucleotide comprising the following nucleic acid sequence:
- A-B-C-D-E
- wherein:
- A is a start codon;
- B is a polynucleotide sequence of 13 nucleic acids, wherein the polynucleotide sequence has homology to SEQ ID NO:2, wherein at least 5 of the 13 nucleic acids are identical to the corresponding position in SEQ ID NO:2, and wherein the nucleic acid sequence does not code for a stop codon;
- C is a polynucleotide having the nucleic acid sequence of SEQ ID NO:20;
- D is the polynucleotide sequence as shown in SEQ ID NO:4;
- and
- E is a polynucleotide sequence selected from the group consisting of:
- a) SEQ ID NO:5;
- b) SEQ ID NO:6;
- c) SEQ ID NO:7;
- d) SEQ ID NO:8;
- e) SEQ ID NO:9;
- f) SEQ ID NO:10;
- g) SEQ ID NO:11;
- h) SEQ ID NO:
- 12;
- and
- i) SEQ ID NO:13,
- wherein at least 50% nucleotides are either adenosine or thymine;
- and
- wherein only E encodes a stop codon.
19 more elements
A purified polynucleotide comprising the following nucleic acid sequence:
- A-B-D-E
- wherein:
- A is a start codon;
- B is a polynucleotide sequence of 13 nucleic acids, wherein the polynucleotide sequence has homology to SEQ ID NO:2, wherein at least 5 of the 13 nucleic acids are identical to the corresponding position in SEQ ID NO:2, and wherein the nucleic acid sequence does not code for a stop codon;
- D is the polynucleotide sequence as shown in SEQ ID NO:4;
- and
- E is a polynucleotide sequence selected from the group consisting of:
- a) SEQ ID NO:5;
- b) SEQ ID NO:6;
- c) SEQ ID NO:7;
- d) SEQ ID NO:8;
- e) SEQ ID NO:9;
- f) SEQ ID NO:10;
- g) SEQ ID NO:11;
- h) SEQ ID NO:
- 12;
- and
- i) SEQ ID NO:
- 13,
- wherein at least 50% nucleotides are either adenosine or thymine;
- and
- wherein only E encodes a stop codon.
19 more elements
A purified polynucleotide selected from the group consisting of:
- a) a polynucleotide sequence as shown in SEQ ID NO:21;
- b) a polynucleotide sequence as shown in SEQ ID NO:22;
- and
- c) a polynucleotide sequence as shown in SEQ ID NO:23.
1 more element
Dependencies are inferred from the claim text.