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US 7,119,187 B2Granted
Internal ribosome entry site of the labial gene for protein expression
AssigneeNational Health Research Institutes
InventorsDung-Fang Lee, Jyh-Lyh Juang
Life of this patent
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Abstract
The invention describes compositions and methods for recombinant protein expression in a wide range of cell types. The compositions comprise an IRES sequence from the Drosophila labial (lab) gene, or a variant or fragment thereof, or alternatively, a homolog of a lab IRES, or a variant or fragment thereof. Methods of using the compositions are also described.
Claims16
A nucleic acid vector for the expression of at least two cistrons comprising:
- a. a promoter operably linked to a nucleotide sequence comprising at least two cistrons;
- and
- b. a nucleotide sequence that provides IRES activity operably linked to each cistron subsequent to the first cistron, wherein at least one of the nucleotide sequences that provide IRES activity comprises a nucleotide sequence chosen from:
- a nucleotide sequence comprising SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–215 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 45–239 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 45–215 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–74 and 187–239 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–74 and 187–215 of SEQ ID NO. 1;
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 1 by substitution of the nucleotides at positions 124–127 of SEQ ID NO. 1;
- a nucleotide sequence comprising SEQ ID NO. 2;
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 2 by substitution of the nucleotides at positions 136–139 of SEQ ID NO. 2;
- and
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 2 by substitution of the nucleotides at positions 126–129 of SEQ ID NO. 2.
11 more elements
A method for expressing at least two cistrons comprising:
- introducing into a host cell a nucleic acid vector comprising:
- a. a promoter operably linked to a nucleotide sequence comprising at least two cistrons;
- and
- b. a nucleotide sequence that provides IRES activity operably linked to each cistron subsequent to the first cistron, wherein at least one of the nucleotide sequences that provide IRES activity comprises a nucleotide sequence chosen from:
- a nucleotide sequence comprising SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–215 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 45–239 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 45–215 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–74 and 187–239 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–74 and 187–215 of SEQ ID NO. 1;
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 1 by substitution of the nucleotides at positions 124–127 of SEQ ID NO. 1;
- a nucleotide sequence comprising SEQ ID NO. 2;
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 2 by substitution of the nucleotides at positions 136–139 of SEQ ID NO. 2;
- and
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 2 by substitution of the nucleotides at positions 126–129 of SEQ ID NO. 2.
12 more elements
A baculovirus transfer vector for the expression of at least two cistrons comprising:
- a. a polyhedrin promoter operably linked to a nucleotide sequence comprising at least two cistrons;
- and
- b. a nucleotide sequence that provides IRES activity operably linked to each cistron subsequent to the first cistron, wherein at least one of the nucleotide sequences that provide IRES activity comprises a nucleotide sequence chosen from:
- a nucleotide sequence comprising SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–215 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 45–239 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 45–215 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–74 and 187–239 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–74 and 187–215 of SEQ ID NO. 1;
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 1 by substitution of the nucleotides at positions 124–127 of SEQ ID NO. 1;
- a nucleotide sequence comprising SEQ ID NO. 2;
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 2 by substitution of the nucleotides at positions 136–139 of SEQ ID NO. 2;
- and
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 2 by substitution of the nucleotides at positions 126–129 of SEQ ID NO. 2.
11 more elements
A recombinant baculovirus capable of expressing at least two cistrons in a host cell comprising a baculovirus genome comprising:
- a. a polyhedrin promoter operably linked to a nucleotide sequence comprising at least two cistrons;
- and
- b. a nucleotide sequence that provides IRES activity operably linked to each cistron subseauent to the first cistron, wherein at least one of the nucleotide sequences that provide IRES activity comprises a nucleotide sequence chosen from:
- a nucleotide sequence comprising SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–215 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 45–239 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 45–215 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–74 and 187–239 of SEQ ID NO. 1;
- a nucleotide sequence comprising nucleotides 1–74 and 187–215 of SEQ ID NO. 1;
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 1 by substitution of the nucleotides at positions 124–127 of SEQ ID NO. 1;
- a nucleotide sequence comprising SEQ ID NO. 2;
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 2 by substitution of the nucleotides at positions 136–139 of SEQ ID NO.2;
- and
- a nucleotide sequence that differs from a nucleotide sequence comprising SEQ ID NO. 2 by substitution of the nucleotides at positions 126–129 of SEQ ID NO. 2.
11 more elements
Dependencies are inferred from the claim text.